WO2021233996A1

WO2021233996A1 - Method and system for fighting pandemic

Info

Publication number: WO2021233996A1
Application number: PCT/EP2021/063287
Authority: WO
Inventors: Prof. Hans LEHRACH; Andele CONRADIE; Bodo Lange; Sebastian Meier-Ewert; Artur MURADYAN
Original assignee: Alacris Theranostics Gmbh
Priority date: 2020-05-19
Filing date: 2021-05-19
Publication date: 2021-11-25

Abstract

The present invention relates to a method and an infrastructure for fighting a pandemic caused by a pathogen by repetitive testing of all humans in specific populations at sufficiently high frequency in order to eliminate specific infectious diseases essentially within a few infection cycles, or collect other medically relevant information. The method is based on collection of body samples, such as saliva, blood from finger-pricks or other relevant types, from an individual at home or at appropriate collection centers using supplied collection tube(s) that can be linked to an individual's infection status, or other medically relevant information and the subsequent processing, amplification by PCR or isothermal amplification and readout of the results either directly or by pooling and massive parallel sequencing of millions of samples using DNA sequence analysis technologies, such as Next Generation Sequencing (NGS). Analysis results are tracked through an appropriate communication device that can identify the infection status of the sample, provide information on necessary quarantine measures to the infected individual, and trigger the notifications on communication devices of other individuals, which have recently been in close proximity to the infected person. The present invention also discloses a method to synthetize molecular labels to identify test tubes for nucleic acid sequence analyses.

Description

Method and system for fighting pandemic

Specification

The present invention relates to a method and an infrastructure for fighting a pandemic caused by a pathogen by repetitive testing of all humans in specific populations at sufficiently high frequency in order to eliminate specific infectious diseases essentially within a few infection cycles, or collect other medically relevant information.

The method is based on collection of body samples, such as saliva, blood from finger-pricks or other relevant types, from an individual at home or at appropriate collection centers using supplied collection tube(s) that can be linked to an individual’s infection status, or other medically relevant information and the subsequent processing, amplification by PCR or isothermal amplification and readout of the results either directly or by pooling and massive parallel sequencing of millions of samples using DNA sequence analysis technologies, such as Next Generation Sequencing (NGS). Analysis results are tracked through an appropriate communication device that can identify the infection status of the sample, provide information on necessary quarantine measures to the infected individual, and trigger the notifications on communication devices of other individuals, which have recently been in close proximity to the infected person.

The present invention also discloses a method to synthetize molecular labels to identify test tubes for nucleic acid sequence analyses.

Background of the invention

The COVID-19 pandemic, now a few months old, has changed the world beyond recognition. As of Mai 2020, the COVID-19 pandemic has already resulted in millions of infected individuals and hundreds of thousands deads, moreover livelihoods have been destroyed, and the global economy is now experiencing its deepest recession in almost a century. Early epidemiological studies indicate that the exponential spread of COVID-19, the disease caused by SARS-CoV-2, can be slowed by restrictive isolation measures, but these measures place an enormous burden on societies and economies. Moreover, once isolation measures are lifted, exponential spread is predicted to resume. The problem is exacerbated by the fact that many infected individuals do not show symptoms, are unlikely to seek testing, and unknowingly contribute to the spread of infection. To effectively combat the transmission of SARS-CoV-2, population-scale testing that can enable rapid identification of all infected individuals is needed, identifying potential outbreak areas to support epidemiological interventions that maximize human health, while reducing the extent of restrictive isolation measures.

The importance of testing has been stressed by many, however, the fact that in a disease with a majority of carriers with no or minimal symptoms the appropriate metric of success is not the fraction tested, but the fraction untested, has maybe not been sufficiently considered

Currently, the majority of SARS-CoV-2 testing is performed using viral RNA extraction followed by RT-qPCR to amplify and detect one of several highly conserved regions of the SARS-CoV-2 genome, or by detecting serum antibodies specific for viral proteins. The global capacity for testing using these approaches, however, has been limited by a combination of access and supply issues for reagents and instruments.

In most cases, nasal and throat swabs are sent to labs, where technicians determine whether a given sample contains the new coronavirus by isolating any viral RNA, turning it into DNA and then using polymerase chain reaction to amplify the DNA to detectable levels. The method has several drawbacks, including the limited number of testing slots, or wells, on a PCR plate, typically 96 or 384, needed to amplify the virus' genetic material.

Analytical methods allowing testing on a massive scale would allow identifying essentially all infected individuals and their close contacts and being able to rapidly deploy quarantine measures in a bid to break the infection chain. Using such approach to interrupt the transfer from all infected individuals, the pathogen could be eliminated from a country within very short time.

The objective of the present invention is solved by the teachings of the independent claims. Further advantageous features, aspects and details of the invention are evident from the dependent claims, the description, and the examples of the present application.

Brief description of the invention

Infectious diseases like the current COVID-19 pandemic can cause enormous human suffering and economic costs in the trillion dollar range due to the restrictive isolation measures, currently the only measure, which has found to be effective in the current epidemic.

Moreover, once these restrictions are lifted, the exponential spread is likely to re- emerge. It has been suggested that population-scale testing can help break the cycle of isolation and spread, but current detection methods are not capable of such large- scale processing.

The lack of population-wide detection methods hampers efforts to accurately assess and stop the spread of SARS-CoV-2. Many scientists are convinced that the big difference between the places that are doing well and the ones that aren't during COVID-19 pandemic are high levels of testing.

The present invention provides here a method and a system for massive parallel testing of all individuals infected with a pathogen identifiable through nucleic acids, i.e. viruses or pathogenic organisms like bacteria, fungi or multicellular pathogens, allowing routine and repeated testing of all humans in specific populations at sufficiently high frequency to be able to eliminate specific infectious diseases within few infection cycles, for example within multiple weeks for SARS-CoV2.

Thus, the present method is aimed to fight a pandemic caused by a pathogen such as SARS-CoV2 in a very efficient and cost-effective way. Indeed, the inventive method allows all the infected individuals to be identified and quarantined in a rapid manner. Moreover, it allows notifying other individuals that they have been in contact with an infected individual. Thus, the infection chain can be stopped in any country and the pathogen, such as SARS-CoV2, can be efficiently and timely eliminated.

Therefore, the inventive method allows avoidance of many deaths, infections, and also the enourmous economic loss linked to a pandemic, to a large extent due to the only current alternative, the longterm application of highly disruptive social distancing measures.

An important advantage of the present invention is to provide analysis of many samples in short time. Indeed, the inventive method allows, in one implementation, through next generation sequencing techniques, to analyse of up to 500 million samples in a single run, allowing keeping the analysis costs very low. Indeed, the actual sequencing costs are estimated at 1 cent per (infected) sample, for DNA fragments of approx. 300 bp, 3000 reads per sample and $10 per Gigabase sequencing cost, or, in another implementation, the analysis of millions of samples per day per (small) processing center by amplification (PCR or isothermal) and endpoint measurement.

Thus, the present application is directed to a method for fighting infectious diseases, and in particular those, which can be transmitted by infected individuals, which can not be easily identified by symptoms, comprising the following steps: mass distribution of sample tubes carrying a "machine-readable identifier" to all individuals within a population and country-wide, for collecting approriate samples, which are typically nasal swabs or saliva samples in the case of SARS-CoV-2; each tube with a machine-readable identifier can also contain oligonucleotides encoding a second, sequence-based barcode, also named "molecular label" to provide a sample-specific unique identifier; the "machine-readable identifier" of the tube containing an individual’s sample is scanned by the individual’s smartphone or a device at the collection center, using an appropriate application, to establish a link between the individual and the sample tube. To establish the identity of the person tested, the test tube and its bar code and an identity document (passport, identity card) can be documented; thereafter, the tube is sent to a designated processing centre for processing; results are sent to the individual via their smartphone or a web based system using e.g. the smartphone- linked unique identifiers and/or the individual, who originally received the sample tube .

The present method is based on population wide collection and processing of body fluid samples (e.g. saliva, gargle fluid, nose or throat swabs etc for SARS-CoV2) in tubes that are linked to individual’s infection status by unique identifiers. The method is, in one implementation, further based on pooling multiple samples, on the scale of millions, and on massive parallel sequencing of DNA fragments providing information on both the individual and the presence of the pathogen.

In a particular embodiment of the invention, in addition to the machine readable barcode, the sample tube already contains oligonucleotides combining two elements: The first element, which functions as sequencing primer, is a nucleic acid sequence serving as a sensor, able to identify the presence and/or relevant features (e.g. antibiotic resistance genes) of the pathogen, by being able to prime on selected regions of the pathogen genome or sequences derived from it. The first element could also be a nucleic acid sequence able to analyse specific host features, for example by being able to prime on selected regions of host genome or transcriptome, specific host features indicating host disease status by priming on or ligating to specific DNA molecules attached to antibodies or aptamers, extension proximity or ligation proximity products.

The second element is a sequence serving as a universal, sequence based identifier, also referred to as "molecular label" either present from the beginning, or assembled from shorter sequences during the construction of the final sequencing templates, linking the individual sample tube (and therefore the individual tested) to the results of the test. This sequence has to be long (and informative) enough to be able to identify a unique sample among all the sample, which might be analysed together (potentially all individuals in a specific country or continent) with sufficient redundancy, that one or more sequencing errors will not lead to a sample misidentification. This can be achieved by synthesizing very many long oligonucleotides (e.g. by ink-jet printer, phpotolitography or similar technologies technologies). The final unique combination of sequences can however alsobe generated by adding specific combinations of shorter oligonucleotides, each from a presynthesised set of similar sequences at different steps along the wayx to construct the final sequencing library, with the combination of sequence specific oligonucleotides use uniquely marking the product of an individual sample. As additional element the sequence can contain a "random nucleic acid sequence stretch" or "unique molecular identifier", to be able to tag the amplification products deriving from a given molecule or insert through each PCR amplification steps. Each "random nucleic acid sequence stretch" is specific of a given molecule or insert of an individual library, and allows avoiding the distorsion of the results due to differential amplification.

In some embodiments, the test tube further comprises a "control sequence" similar to but distinguishable from the pathogen serving as internal control to make more reliable estimates of the number of pathogens per body sample.

In some embodiments, the first steps to generate DNA molecules for sequencing can be carried out by the tested individual following detailed instructions provided with the kits and/or the internet. In some embodiments, the entire process can be carried out at the analysis centres.

The results of the sequencing analyses are stored in a database in an anonymized form, and can be retrieved by the individuals using a smartphone or web-based application that links an individual with their unique identifiers.

The machine-readable identifiers (e.g. barcodes) characterising the individual samples positive for the pathogen are then made available through a special online database allowing individuals to retrieve the results of the test via a smartphone based application. Alternatively, the individuals can be called or notified through other mechanisms.

Thus, the inventive method follows all legal and publicly expected data protection rules by storing the analysis of the body sample in at least one database in an anonymized form, and retriving the analyses of a particular individual on the basis of the identification label connected to the molecular barcode.

In some embodiments, mechanisms such as for example those bluetooth based, can be triggered, notifying other individuals, which preferably have also been subject to the analysis that they have been in close proximity to an infected individual.

In some embodiments, each individual receves one or more test tubes at regulat intervals, where they put their bodily fluid sample at specific intervals (e.g. every day, every two days, twice a week, weekly) and send back the sample tube, in order to follow the development of pathogen infection.

In some embodiments, the infected individuals receive one set of sample tubes that can deliver the total sequencing of the virus genome and information about genes in patients, that are relevant for the immune response, or the total sequencing of the genome or exome of the infected individual.

It has been shown in a clinical study that tests letting the patients swab the front part of their nostril and mid-nose are just as accurate as those given by medical professionals. This is very critical as the quality of testing also will impact the outcome of the result and shows that it is very likely that self testing will be feasible in conjunction with mass testing ( Tu YP, et al., medRxiv 2020. Patient-collected tongue, nasal, and mid-turbinate swabs for SARS-CoV-2 yield equivalent sensitivity to health care worker collected nasopharyngeal swabs). Similarly positive results have also be achieved using saliva or gargle samples, also simplifying self-testing.

Detailed description of the invention

The present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, or appropriate subgroups, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing or end point measurements on PCR products or products of isothermal amplification strategies, preferably by end-point PCR or equivalent amplification techniques, or nucleic acid sequencing; e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based system in a database. h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products or i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, or n3^') granting the individual access to the at least one database through a web based interface, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Preferred, a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed sample to one of the analysis laboratories; or h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Alterntaively, instead of n3) step, n3^') is performed as follows: n3^') granting the individual access to the at least one database through a web based interface, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Preferred, a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by end-point PCR or equivalene amplification techniques, or nucleic acid sequencing; e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based system in a database. h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

In particular, the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; i) pooling of a plurality of processed samples collected in the test tubes;

Alterntatively, instead of n3) step, n3^') is performed as follows: n3^') granting the individual access to the at least one database through a web based interface, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

In another embodiment, the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, individual, or through a web based interface, in the database; h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products or i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label. Alterntaively, instead of n3) step, n3^') is performed as follows: n3^') granting the individual access to the at least one database through a web based interface, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Moreover, the present invention relates to a method for fighting a pandemic caused by a pathogen by repetitive testing of essentially all individuals in specific populations at sufficiently high frequency in order to eliminate specific infectious diseases essentially within a few infection cycles, and to carry out cost-effectively a wide range of other tests important for individualising therapy, prevention or well-being of many individuals.

Repetitive testing of all individuals of a population comprises repeating the steps f) - p) of the method after a period comprised between 1 week to one year, preferably between 1 to 3 weeks, until when all individual of a population result negative for the pathogen causing the infectious diseases.

Thus, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products or i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, q) repeating steps f) - n) of the method after a period comprised between 1 week to one year, one day to 3 weeks, preferably between one day to 1 week, until when all individuals of the population result negative for the pathogen causing the infectious diseases.

Optionally, in the step h^') the processing further comprises directly determing the concentration of the specfic amplicaton produts, thus h^') may read as follows: h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products. Alterntaively, instead of n3) step, n3^') is performed as follows: n3^') granting the individual access to the at least one database through a web based interface, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Moreover, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes; L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, o)requesting an individual resulted positive for the pathogen causing the infectious disease to follow an isolation period and to follow the recommendations for the treatment of the infectious disease; p) notifying other individuals that they have been in proximity to an individual resulted positive for the pathogen causing the infectious disease, and requesting these other individuals to follow a quarantine period; q) repeating steps f) - p) of the method after a period comprised between 1 week to one year, one day to 3 weeks, preferably between one day to 1 week, until when all individuals of the population result negative for the pathogen causing the infectious diseases.

Optionally, in the step h^') the processing further comprises directly determing the concentration of the specfic amplicaton produts, thus h^') may read as follows: h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products.

Sample tracking

After the analysis of the collected body samples, the individuals can be contacted through different systems: a) A top-down system, in which a central organisation keeps track of the specific sample tube through the identification label that has been provided to an individual. This makes it relatively straightforward to contact the infected individuals, to rule out further infection through quarantine, and eventually to provide information to contact tracing applications. b) A bottom-up approach, centred on the link between the sample identification label and the individual communication device (i.e. a smartphone), established by scanning the identification label of the sample tubes before sending them to analysis laboratories. Through an appropriate application, the smartphone can identify the infection status of the sample, provide information on necessary quarantine measures to the infected individual. This option would help to avoid data protection issues. c) The identification labels (e.g. barcodes) characterising the individual body samples resulted positive or negative for the pathogen are made available through an online database allowing individuals to get the results of the test.

A "communication device" is a hardware device capable of transmitting an analog or digital signal over the telephone, other communication wire, or wirelessly.

Examples of communication devices are bluetooth devices, smartphones, any portable, mobile, hand-held or miniature consumer electronic device, cellular phones, other wireless communication devices, personal digital assistances, programmable remote controls, pagers, laptop computers, tablets, a smartwatch, a personal digital assistant and/or an e-reader device.

"Collection centers" include, but are not limited to, test centers, medical facilities, medical offices, medical laboratories, pharmacies, doctors, supermarkets, gas stations etc.which collect the the test tubes from essentially each individual of a population personally or by mail.

In some embodiments of the present disclosure, a medical professional may obtain a biological sample for testing. In some cases the medical professional may refer the subject to a testing center or laboratory for submission of the biological sample. In other cases, the subject may provide the sample.

The analysis laboratory may enter body sample information into a "database" following an anonymization or pseudonymization procedure for the purpose of one or more of the following: inventory tracking, assay result tracking, order tracking, individual management, individual service, and others. "Pseudonymization" is a data management and de-identification procedure by which personally identifiable information fields within a data record are replaced by one or more artificial identifiers, or pseudonyms, linked to the unique identification label. A single pseudonym for each replaced field or collection of replaced fields makes the data record less identifiable while remaining suitable for data analysis and data processing.

Body sample information entered in the database may include, but is not limited to: individual name, unique individual identification label linked to the test tube, individual associated medical professional, analysis results, adequacy status, indicated adequacy tests, medical history of the individual, preliminary diagnosis, suspected diagnosis, sample history, insurance provider, medical provider, third party testing center or any information suitable for storage in a database. Body sample history may include but is not limited to: age of the sample, type of sample, method of acquisition, method of storage, or method of transport.

The database may be accessible by an individual, medical professional, third party, or any individual or entity which the analysis laboratory grants access. Database access may take the form of electronic communication such as through a communication device such as a smartphone, a computer or telephone. The database may be accessed through an intermediary such as a consultant, independent testing center, or medical professional. The degree of database access or sample information is restricted to comply with generally accepted or legal requirements for patient or customer confidentiality.

The inventive method comprises the following steps:

1 ) Collection of body samples

2) Sample processing: b) cDNA synthesis by reverse transcriptase and virus specific and, for sequencing analysis, sample specific primers /primer combinations c) Target amplification: amplification of the virus genome present in the cDNA using PCR or other i(isothermal) amplification reactions and virus specific and for sequencing analysis, sample specific primers/primer combinations d’) direct determination of the concentration of the specific amplification products by fluorescent readers or d”) pooling of amplification products from different individuals) Library preparation: fragmentation and addition of adapter sequences (tagging) and evtl. unique molecular identifier

3) Pooling of different libraries from different pools of individuals

4) analysis of pooled samples

5) data storing

6) notification of the results

Collection of body samples

Herein, a "body sample" is a sample of bodily fluid, e.g. from an individual which is to be subjected to the herein disclosed method. Such body samples may be from a human, a plant or an animal. Preferred samples include saliva, gargle lavage, blood from finger-prick, nasal swab, oropharyngeal swab, tongue swab, mid-turbinate swab, throat swab, nasal wash, sputum, blood, serum, plasma, cerebrospinal fluid, urine, and pleural effusions. Preferably, the sample is a saliva sample, a throat swab sample, or a gargle lavage sample, a sputum sample, or a nasal swab, more preferably, the sample is a saliva sample, a sputum sample, or a nasal swab, most preferably a saliva sample, a throat or nose swab sample, a gargle lavage sample, or a sputum sample.

As previously shown, tongue, nasal, and mid-turbinate swab samples self-collected from the patients give similar results to those collected by medical professional ( Tu YP, et al., medRxiv 2020. Patient-collected tongue, nasal, and mid-turbinate swabs for SARS-CoV-2 yield equivalent sensitivity to health care worker collected nasopharyngeal swabs).

A gargle laveage sarnie is also self-collected from the patients gives similar results (David M. Goldfarb et al, Journal of Clinical Microbiology, 2021 , 59(4), e02427-20).

Moreover, each individual can collect more than one body sample, each body sample in one test tube, in order to analyse body samples of different origin, or to perform the analysis in replicate.

The test tube comprises a cap containing PSMF, a layer containing proteinase K, a layer consisting of wax, a master mix for a RT-PCR reaction, and a combination of individuum specific primer.

Thus, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a saliva sample, a blood sample from finger-prick, a sputum sample, a gargle lavage sample, a tongue swab, a throat swab, or a nasal swab, preferably a gargle laveage sample or a nasal swab, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

Thus, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a saliva sample, a blood sample from finger-prick, a sputum sample, a gargle lavage sample, a tongue swab, a throat swab, or a nasal swab, preferably a gargle laveage sample or a nasal swab, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, or wherein processing the samples either before or after shipment comprises linking or amplifying a molecular label sequence uniquely indicating the original test tube.

The "test tube" or "sample tube" or "sample container" may be any container suitable for storage and or transport of the body sample including but not limited to: a cup, a cup with a lid, a tube, a sterile tube, a vacuum tube, a syringe, a bottle, a microscope slide, or any other suitable container. The test tube is preferably sterile. The test tube comprises an identification label, a molecular label and a preservation material, to ensure the stability of the sample and precision of the analysis after the delivery and eventually storage of the sample. An examplary preservation material is ethanol.

Moreover, each individual can receive more than one test tube to collect body samples of different origin, or to perform the analysis in replicate.

Processing of the samples

In some embodiments, "processing of the body sample" also comprises body sample lysis, virus lysis, nucleic acid extraction, eventual cDNA transcription in the cases of RNA-based pathogens, and amplification with pathogen specific oligonucleotides, in order to amplify pathogen specific nucleic acid sequences.

In some embodiments, "processing of the body sample" further comprises DNA fragmentation and addition of adapter sequences (tagging), and eventually of unique molecular identifier, to prepare the individual library.

In some embodiments, "processing of the body sample" can also comprise treatment with proteinase K in order to inactivate RNase enzymes and protect RNA.

The wording "pooling of a plurality of samples" or "pooling of a plurality of processed samples" refers to the pooling of the samples collected by more individuals in order to analyse many samples in a single run. In some embodiments, this disclosure provides for pooling of samples, such as of 10² - 10⁵ samples, 10² - 10⁶ samples, 10² - 10⁷ samples, 10² - 10⁸ samples,

10² - 10⁹ samples, 10² - 10¹⁰ samples, 10² - 10¹¹ samples, 10² - 10¹² samples,

5 x 10² - 5 x 10¹² samples, 10³ — 10⁵ samples, 10³ — 10⁶ samples, 10³ — 10⁷ samples,

10³ — 10⁸ samples, 10³ - 10⁹ samples, 10³ - 10¹⁰ samples, 10³ - 10¹¹ samples,

10³ - 10¹² samples, 5 x 10³ - 5 x 10¹² samples, 10⁴ - 10⁵ samples, 10⁴ - 10⁶ samples, 10⁴ - 10⁷ samples, 10⁴ - 10⁸ samples, 10⁴ - 10⁹ samples, 10⁴ - 10¹⁰ samples, 10⁴ - 10¹¹ samples, 10⁴ - 10¹² samples, 5 x 10⁴ - 5 x 10¹² samples,

10⁵ - 10⁶ samples, 10⁵ - 10⁷ samples, 10⁵ - 10⁸ samples, 10⁵ - 10⁹ samples, 10⁵ - 10¹⁰ samples, 10⁵ - 10¹¹ samples, 10⁵ - 10¹² samples, 5 x 10⁵ - 5 x 10¹² samples, 10⁶ - 10⁷ samples, 10⁶ - 10⁸ samples, 10⁶ - 10⁹ samples, 10⁶ - 10¹⁰ samples, 10⁶ - 10¹¹ samples, 10⁶ - 10¹² samples, 5 x 10⁶ - 5 x 10¹² samples,

10⁷ — 10⁸ samples, 10⁷ - 10⁹ samples, 10⁷ - 10¹⁰ samples, 10⁷ - 10¹¹ samples, 10⁷ - 10¹² samples, 5 x 10⁷ - 5 x 10¹² samples, 10⁸ - 10⁹ samples, 10⁸ - 10¹⁰ samples, 10⁸ - 10¹¹ samples, 10⁸ - 10¹² samples, 5 x 10⁸ - 5 x 10¹² samples, 10⁹ - 10¹⁰ samples, 10⁹ - 10¹¹ samples, 10⁹ - 10¹² samples, 5 x 10⁹ - 5 x 10¹² samples, 10¹⁰ - 10¹¹ samples, 10¹⁰ - 10¹² samples, 5 x 10¹° - 5 x 10¹² samples.

The number of samples that can be pooled depends on the length of molecular label. If the molecular label is 5 bases, up to 1000 samples can be pooled, if the molecular label is 10 bases, up to 10⁶ samples can be pooled, if the molecular label is 15 bases, up to 10¹² samples can be pooled.

An identification label refers to a machine readable identifier, for example a barcode. Therefore, "identification label", "machine readable identifier", and "machine readable identification label" can be used interchangeably.

A "molecular label" refers to one or more nucleic acid sequences present in the test tube before sample collection, or added during sample processing informationally linked to the "machine readable-identifier" and thus, to the result of the test, and which will be sequenced in the final analysis of the pooled samples. The molecular label allows distinguishing between billions of individual body samples.

In some embodiments, the molecular label is comprised in the oligonucleotide complementary to a nucleic acid sequence specific for the pathogen, which is used prime and amplify said nucleic acid sequence.

In alternative embodiments, the "molecular label" can be assembled from shorter sequences during analysis and in particular during the construction of the final sequencing templates. In other words, the "molecular label" can be assembled by amplifying nucleic acid sequences (for example nucleic acid sequences from genome regions at high variability) of the individual. The presence of this tag or of specific sequence elements in the final template allows to identify the test results of a specific individual.

"Molecular labels" useful in the method described herein are less than or equal to 50, or 40, or 30, or 20, or 15, or 10, or 7 but preferably longer than 10 nucleotides in length.

As additional elements, the sequence can contain a "random nucleic acid sequence stretch" or "unique molecular identifier", to be able to tag the products of every single molecule through amplification steps, as well as appropriate control sequences as an aid in normalising the results of the analysis.

A "random nucleic acid sequence stretch" useful in the method described herein is less than or equal to less than or equal to 50, or 40, or 30, or 20, or 15, or 10, or 7 but preferably longer than 10 nucleotides in length.

Thus, the "unique molecular identifier" ("UMI") are useful to avoid the distorsion of the results through differential amplification during the target amplification. The "unique molecular identifier" are short, random sequences comprised inside the used primer, that can mark each molecule with their own sequence, and in this way allow to follow the amplification products of the same molecules.

The link between one molecular label comprising at least one nucleic acid sequence and an identification label can occur in different ways.

The molecular identifier can be synthesised separately, linked to the machine readable identifier, e.g. by synthesising the nucleic acid sequences containing the unique identifiers on spots on a substrate (plastic band etc), also marked with a unique machine readable bar code, cutting out the spots and transferring them to the sample tube,

The sequence of the randomly synthesised molecular identifier can be correlated with the machine readable identifier in a combinatorial synthesis by reading electronically the machine readable identifiers passing through the individual (random) synthesis step.

In theory, the sequences of the oligonucleotides in all sample tubes could also be determined by sequencing a small sample. All these strategies are however not straightforward, due to the need for sufficiently long unique identifiers as part of the final construct.

To obviate the need for synthesising millions or billions unique long nucleotides, and to increase the range of tests, which can be carried out in the same sample in parallel, we describe here a modified strategy essentially replacing the one molecule carrying all the information by a sequence library based on three identifiers:

- the barcode on the tube

- the unique (but unknown) sequence of the bead oligo

- the known molecular label of the tube established by copying information from a combination of oligonucleotides, selected in such a way to uniquely define the tube

A molecular label identifying a test tube can by produced by constructing beads carrying DNA sequences consisting of the following elements (5’ to 3’): a common sequence to be used later to introduce sequence elements required for the sequencing process (library arms), by deterministic synthesis, a sequence identifying each bead by combinatorial synthesis (in each step, all beads are randomly distributed to reaction vessels extending the growing chain by one or more bases) potentially a random sequence differing between different molecules (by synthesis with a mixture of all 4 bases in each step) an adaptor sequence, used later to introduce different sequence elements to carry out specific functions linked to the unique identifier for the bead.

To be able to introduce multiple functions into the sequences of bead, enzymatic techniques (ligation, site specific recombinases etc can be used). At least two functional elements are introduced: one or more sequences needed for the specific test or tests to be carried out (e.g. Corona virus genome, Influenza genome, host alleles etc) one or more sequences required to ultimately provide a (known) identifier of the sequence.

Thus, a preferred embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label, or wherein a molecular label identifying the test tube is assembled at step h) or h') with nucleic acid sequences of the individual, and wherein analyzing the pooled processed samples at step I) further comprises nucleic acid sequencing of said molecular label.

Preferably, in the step a) the body sample is a saliva sample, a blood sample from finger-prick, a sputum sample, a gargle lavage sample, a tongue swab, a throat swab, or a nasal swab, more preferably a gargle laveage sample or a nasal swab.

Reverse transcription and product amplification

Amplification of viral DNA segments is important to increase the probability of virus detection, and to compensate the material losts intrinsic in the procedure.

Thus, preferred, the step I) comprises amplication of viral DNA segment, when a body sample containing viral RNA or DNA.

Suitable product amplification procedures comprise: polymerase chain reaction (PCR), Recombinase Polymerase Amplification (RPA), RRA, helicase-dependent amplification (HDA), Loop-mediated isothermal amplification (LAMP), etc. Suitable product amplification procedures can be combined with the cDNA transcription, such as in: RT-PCR, endpoint PCR, RT-RPA, RT-RRA, RT-HDA, RT-LAMP. Preferred, endpoint PCR (ePCR) is applied.

Thus, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a saliva sample, a blood sample from finger-prick, a sputum sample, a gargle lavage sample, a tongue swab, a throat swab, or a nasal swab; preferably a gargle laveage sample or a tnasal swab; b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes,

I) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by either nucleic acid sequencing or an amplification-end-point detection method selected from RT-PCR, RT-RPA, RT-RRA, RT-HDA, and RT-LAMP; at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Thus, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a saliva sample, a blood sample from finger-prick, a sputum sample; a gargle lavage sample, a tongue swab, a throat swab, or a nasal swab, preferably a gargle laveage sample or a nasal swab; b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing or an amplification-end-point detection method based on RT-PCR, RT- RPA, RT-RRA, RT-HDA, and RT-LAMP at the analysis laboratory,; m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label wherein processing the samples either before or after shipment comprises linking or amplifying a molecular label sequence uniquely indicating the original test tube.

More preferred, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a gargle laveage sample or a nasal swab; b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes,

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease either by nucleic acid sequencing or by endpoint PCR; or an equivalentend-point amplification detection (RPA etc) at the analysis laboratory; m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

Still more preferred, the present invention is also directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, wherein the body sample is a gargle laveage sample or a nasal swab; b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, and shipping the processed body sample to one of the analysis laboratories; or h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

I) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by either nucleic acid sequencing or by endpoint PCR or an equivalentend-point amplification detection (RBA etc) at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein processing the samples either before or after shipment comprises linking or amplifying a molecular label sequence uniquely indicating the original test tube.

NUCLEIC ACID SEQUENCING

The nucleic acid sequencing tecnique can be a next generation sequencing technique. For example, next generation sequencing technique may be sequencing by synthesis, pyrosequencing, sequencing by oligo ligation, semiconductor technology or single molecule real-time (SMRT™) sequencing. However, the herein disclosed method is not restricted to large scale sequencing methods. Local sequencing methods (e.g. Nanopore Sequencers in USB stick format) could be part of large scale testing efforts as well.

Specific examples of sequencing techniques are: (i) Sequencing by synthesis chemistry: lllumina: HiSeq 2000, HiSeq 1000, Genome Analyzerl lx, MiSeq, HiScanSQ,

(ii) Pyrosequencing chemistry: Roche 454 GS FLX, GS FLX+, GS Junior,

(iii) Sequencing by oligo ligation chemistry: Invitrogen: SOLD 5500 Series,

(iv) Semiconductor technology chemistry: Invitrogen: lonTorrent PGM or Ion Proton Sequencer,

(v) Single molecule, real-time (SMRT™) sequencing chemistry: Pacific Biosciences, PacBio RS system

(vi) Oxford Nanopore sequencing

Any standard next-generation technique can be used to perform the invention described herein, as for example those described in " Next Generation Sequencing, Methods and Protocols, Editors: Steven R., Ordoukhanian P., Daniel R.".

The introduction of targeting elements to the bead identifier sequence is preferably done by a method selected from the group of assembly by overlap extension polymerase chain reaction, site-specific recombination, and/or ligation.

In the context of the present invention the target "nucleic acid sequence specific to the pathogen" or "of the individuals" may be, inter alia, RNA, DNA, cDNA (complementary DNA), mRNA (messenger RNA), mtRNA (mitochondrial RNA), rRNA (ribosomal RNA), tRNA (transfer RNA), nRNA (nuclear RNA), siRNA (short interfering RNA), snRNA (small nuclear RNA), snoRNA (small nucleolar RNA), scaRNA (Small Cajal Body specific RNA), microRNA, dsRNA (doubled-stranded RNA), ribozyme, riboswitch, viral RNA, dsDNA (double-stranded DNA), ssDNA (single-stranded DNA), plasmid DNA, cosmid DNA, chromosomal DNA, viral DNA, mtDNA (mitochondrial DNA), nDNA (nuclear DNA) and snDNA (small nuclear DNA). Preferably however, the target nucleic acid sequence specific the pathogen or of the individuals are cDNA (complementary DNA), mRNA (messenger RNA), mtRNA (mitochondrial RNA), nRNA (nuclear RNA), microRNA, dsRNA (doubled-stranded RNA), viral RNA, dsDNA (double-stranded DNA), ssDNA (singlestranded DNA), plasmid DNA, chromosomal DNA, viral DNA, mtDNA (mitochondrial DNA), nDNA (nuclear DNA). More preferably, the target nucleic acid molecules are viral RNA.

As used herein, a "polynucleotide" or "nucleic acid sequence" or "oligonucleotide" generally refers to any "polynucleotide" or "nucleic acid sequence" or "oligonucleotide", which may be unmodified RNA or DNA or modified RNA or DNA. "Nucleic acid sequence" includes, without limitation, single- and double- stranded nucleic acid sequences. The term "nucleic acid sequence" as it is used herein embraces chemically, enzymatically or metabolically modified forms of nucleic acid sequence, as well as the chemical forms of DNA and RNA characteristic of viruses and cells, including for example, simple and complex cells.

As used herein, the term "cDNA" refers to complementary or copy oligonucleotide produced from an RNA template by the action of an RNA-dependent DNA polymerase activity (e.g., reverse transcriptase).

As used herein, "complementary" refers to the ability of a single strand of an oligonucleotide (or portion thereof) to hybridize to an anti-parallel oligonucleotide strand (or portion thereof) by contiguous base-pairing between the nucleotides (that is not interrupted by any unpaired nucleotides) of the anti-parallel oligonucleotide single strands, thereby forming a double-stranded oligonucleotide between the complementary strands. A first oligonucleotide is said to be "completely complementary" to a second oligonucleotide strand if each and every nucleotide of the first oligonucleotide forms base-paring with nucleotides within the complementary region of the second oligonucleotide. A first oligonucleotide is not completely complementary (i.e. , partially complementary) to the second oligonucleotide if one nucleotide in the first oligonucleotide does not base pair with the corresponding nucleotide in the second oligonucleotide. The degree of complementarity between oligonucleotide strands has significant effects on the efficiency and strength of annealing or hybridization between oligonucleotide strands. This is of particular importance in amplification reactions, which depend upon binding between oligonucleotide strands.

An oligonucleotide primer is "complementary" to a target polynucleotide or nucleic acid sequence if at least 50% (preferably, 60%, more preferably 70%, 80%, still more preferably 90% or more) nucleotides of the primer form base-pairs with nucleotides on the target nucleic acid sequence.

As used herein, the term "detect" or "detection" refers to the qualitative determination of the presence or absence of a pathogen in a body sample. The term "detect" or "detection" also includes the "identification" of the pathogen, i.e., determining the genus, species, or strain of the pathogen according to recognized taxonomy in the art. The term "detect" or "detection" further includes the quantitation of a microorganism in a sample, e.g., the copy number of the microorganism in a microliter (or a milliter or a liter) or a microgram (or a milligram or a gram or a kilogram) of a sample.

As used herein, a "nucleic acid sequence specific for the pathogen" is a target nucleic acid sequence or polynucleotide as defined above, which is present in only one pathogen that is being analyzed. As used herein, an "oligonucleotide or oligonucleotide primer complementary to at least one nucleic acid sequence specific for the pathogen" refers to a polynucleotide molecule (i.e., DNA or RNA) capable of annealing to a polynucleotide template and providing a 3' end to produce an extension product that is complementary to the polynucleotide template. The conditions for initiation and extension usually include the presence of four different deoxyribonucleoside triphosphates (dNTPs) and a polymerization-inducing agent such as a DNA polymerase or reverse transcriptase activity, in a suitable buffer ("buffer" includes substituents which are cofactors, or which affect pH, ionic strength, etc.) and at a suitable temperature. The oligonucleotide primer as described herein may be single- or double-stranded. The oligonucleotide primer is preferably single-stranded for maximum efficiency in amplification. "Oligonucleotide" useful in the methods described herein are less than or equal to 100 nucleotides in length, e.g., less than or equal to 90, or 80, or 70, or 60, or 50, or 40, or 30, or 20, or 15, but preferably longer than 10 nucleotides in length.

Moreover, the pathogen genome sequencing can be combined with other analyses, such as:

- pathogen proteins can be detected by an assay such as Proximity Extension Assay (PEA), which is an immunoassay for high throughput detection of protein biomarkers in liquid samples. For each biomarker, a matched pair of antibodies linked to unique oligonucleotides (proximity probes) binds to the respective protein target. As a result, the probes come in close proximity and hybridize to each other. The addition of a DNA polymerase leads to an extension of the hybridizing oligo, bound to one of the probes, creating a DNA amplicon that can subsequently be detected and quantified by quantitative real-time PC or by sequencing.

- detection of nucleic acid modification (methylation) at specific sites or on a subgenic basis

Thus, a more preferred embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one machine readable identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, and wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label, or wherein a molecular label identifying the test tube is assembled at step h) or h') with nucleic acid sequences of the individual, and wherein analyzing the pooled processed samples at step I) further comprises nucleic acid sequencing of said molecular label, and wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify the at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence.

Optionally, the step I) comprises amplication of viral DNA segment, when a body sample contains viral RNA or DNA. Preferably, the amplication of viral DNA segment is RT-PCR or for larger sample pools more preferably e-PCR.

Moreover, a still more preferred embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of an individual, b) providing one machine readable identification label for each test tube, c) optionally generating or providing a molecular label comprising a nucleic acid sequence; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) optionally processing the sample, and shipping the test tube to one of the analysis laboratories; or h’) shipping the test tube to one of the analysis laboratories and optionally processing the sample; i) pooling of a plurality of processed samples collected in the test tubes; j) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, k) storing the analysis of the processed sample in the at least one database in an anonymized form,

L) notifying the analysis of the processed sample to the individual by:

11 ) notifying the analysis to said individual identified through the identification label by contacting said individual;

12) notifying the analysis to said individual through the communication device of step g) identified through the identification label;

13) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify the at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence; wherein processing the sample comprises connecting a molecular label comprising a nucleic acid sequence to a nucleic acid sequence indicating the result of the test; and/or optionally generating a unique identifier sequence linked to the machine readable identification label of the test tube.

Optionally, the step I) comprises amplication of viral DNA segment, when a body sample containing viral RNA or DNA. Preferably, the amplication of viral DNA segment is RT-PCR or for larger sample pools, e-PCR, more preferably e-PCR.

REFERENCE CONTROL

In some embodiments, the test tube further comprises a pathogen reference nucleic acid sequence distinguishable by its sequence in the final test result.

The pathogen reference nucleic acid sequence is present at specific concentration and serves as internal control to make more reliable estimates of the number of pathogens per body sample.

As used herein the "reference nucleic acid sequence specific for the pathogen" is preferably a synthetic RNA or DNA spike-in that contains same priming regions as the target nucleic acid sequence specific for the pathogen we are trying to detect, but the amplicon has a unique string of nucleotides that allows to differentiate it from the actual pathogen sequence by sequencing. This reference nucleic acid sequence serves as a well control for amplification of the virus sequence, but also as a way to allow for amplification even in negative samples and thus providing normalization between samples.

Therefore, according to one embodiment, the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

According to a preferred embodiment, the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, and wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify the at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence, and wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

Optionally, in the step h^') the processing further comprises directly determing the concentration of the specfic amplicaton produts, thus h^') may read as follows: h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products.

According to a more preferred embodiment, the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, and wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label, or wherein a molecular label identifying the test tube is assembled at step h) or h') with nucleic acid sequences of the individual, and wherein analyzing the pooled processed samples at step I) further comprises nucleic acid sequencing of said molecular label, and wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify the at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence, and wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

ANALYSING INDIVIDUAL IMMUNE RESPONSE

The present invention also allows analysing the immune status of the individual. This is achieved by sequencing nucleic acid relevant for the immune response of the individual, such as the HLA alleles, or the VH and VL chains of the antibodies.

Therefore, one embodiment of the present invention further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, and thus comprises sequencing at least one individual nucleic acid sequence relevant for the immune response, in order to assess the immune status of the individual.

"Oligonucleotides" useful in the methods described herein are less than or equal to 100 nucleotides in length, e.g., less than or equal to 90, or 80, or 70, or 60, or 50, or 40, or 30, or 20, or 15, but preferably longer than 10 nucleotides in length.

Therefore, one embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, and wherein the test tube further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, and wherein analysing at step i) the body sample collected in the test tube further comprises sequencing at least one nucleic acid sequence of the individual relevant for the immune response.

Preferably, the at least one individual nucleic acid sequence relevant for the immune response is an immunoglobulin nucleic acid sequence, more preferably is an immunoglobulin VH nucleic acid sequence and an immunoglobulin VL sequence.

Therefore, one more particular embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, and wherein analysing at step I) the body sample collected in the test tube further comprises sequencing at least one nucleic acid sequence of the individual relevant for the immune response, and wherein the test tube further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, and wherein the at least one nucleic acid sequence relevant for the immune response is an immunoglobulin nucleic acid sequence, more preferably is an immunoglobulin VH nucleic acid sequence and an immunoglobulin VL nucleic acid sequence. Optionally, in the step h^') the processing further comprises directly determing the concentration of the specfic amplicaton produts, thus h^') may read as follows: h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products.

Herein, immunoglobulin VH refers to the variable region of the heavy chain of the immunoglobulin, and immunoglobulin VL refers to the variable region of the light chain of immunoglobulins

Moreover, the present method allows measuring other markers of the individuals such as:

- nucleic acid tags on antibodies or aptamers directed against specific proteins or other features, used to either indicate the presence of specific antigens directly, or on the basis of e.g. proximity ligation or proximity extension assays

- nucleic acid tags on other functionally important biological entities

- nucleic acid modification (e.g. methylation)

INFECTIOUS DISEASES. ISOLATION AND QUARANTINE "Communicable diseases", also known as "infectious diseases" or "transmissible diseases", are illnesses that result from the infection, presence and growth of a pathogen or pathogenic biologic agents, capable of causing disease, in an individual human or other animal host. Infections may range in severity from asymptomatic, i.e. without symptoms, to severe and fatal. The term infection does not have the same meaning as infectious disease because some infections do not cause illness in a host.

Disease causing pathogens include viruses, bacteria, fungi, protozoa, multicellular parasites, and aberrant proteins known as prions (potentially detectable through proximity extension type assays). Transmission of these biologic agents can occur in a variety of ways, including direct physical contact with an infectious person, consuming contaminated foods or beverages, contact with contaminated body fluids, contact with contaminated inanimate objects, airborne (inhalation), or being bitten by an infected insect or tick. Some disease agents can be transmitted from animals to humans, and some of these agents can be transmitted in more than one way.

In the Commission Decision No 2000/96/EC of the European Union, the following diseases have been defined as communicable or infection diseases: Anthrax, Botulism, Brucellosis, Campylobacter enteritis, Chikungunya virus disease, Chlamydial infection, including Chlamydial lymphogranuloma (venereum) (LGV), Cholera, Creutzfeldt-Jakob disease, Cryptosporidiosis, Dengue, Diphtheria, Echinococcosis, Giardiasis (lambliasis), Gonococcal infection, Haemophilus influenzae infection, invasive disease, Acute hepatitis A, Hepatitis B, Hepatitis C, Human immunodeficiency virus (HIV) infection and Acquired immunodeficiency syndrome (AIDS), Influenza, Influenza A/H5N1 , Legionnaires' disease, Leptospirosis, Listeriosis, Lyme neuroborreliosis, Malaria, Measles, Meningococcal infection, invasive disease, Mumps, Pertussis, Plague, Streptococcus pneumoniae infection, invasive disease, Acute poliomyelitis, Q fever, Rabies, Rubella, Congenital rubella syndrome, Salmonella enteritis, Severe acute respiratory syndrome (SARS), Shiga toxin/verocytotoxin-producing E. coli infection (STEC/VTEC), including Haemolytic- uraemic syndrome (HUS), Shigellosis, Smallpox, Syphilis, Congenital syphilis, Tetanus, Tick-borne viral encephalitis, Congenital toxoplasmosis, Trichinellosis, Tuberculosis, Tularaemia, Typhoid and paratyphoid fevers, Viral haemorrhagic fevers (VHF), West Nile virus infection, Yellow fever, Enteritis due to Yersinia enterocolitica or Yersinia pseudotuberculosis, Zika virus disease, Congenital Zika virus disease

Isolation and quarantine are procedures helping to protect a population by preventing exposure to individuals who have or may have an infectious disease.

The term "isolation" relates to procedure separating sick people with a quarantinable communicable disease from individuals who are not sick. It typically occurs in a hospital setting, but can be done at home or in a special facility. Usually individuals are isolated, but the practice may be applied in larger groups. Isolation is prolonged until when the individual is not sick anymore and results negative for the pathogen.

The term "quarantine" relates to procedure separating and restricting the movement of people who were exposed to a contagious disease to see if they become sick, i.e. before it is known whether they will become ill. Quarantine usually takes place in the home and may be applied at the individual level or to a group or community of exposed persons. Its maximum duration would be one incubation period from the last known exposure, but it could be shortened if an effective vaccination or prophylactic treatment is available and can be delivered in a timely fashion.

In the United States, isolation and quarantine are authorized for the following infectious diseases: cholera, diphtheria, infectious tuberculosis, plague, smallpox, yellow fever, viral hemorrhagic fevers, severe acute respiratory syndromes, flu that can cause a pandemic, such as for example COVID-19.

Therefore, some embodiments of the present method comprise after step n), the following step o) and p): o) requesting an individual resulted positive for the pathogen causing the infectious disease to follow an isolation period and to follow the recommendations for the treatment of the infectious disease; p) notifying other individuals that they have been in proximity to an individual resulted positive for the pathogen causing the infectious disease, and requesting these other individuals to follow a quarantine period.

Thus, a further embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein process ing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label; o) requesting an individual resulted positive for the pathogen causing the infectious disease to follow an isolation period and to follow the recommendations for the treatment of the infectious disease; p) notifying other individuals that they have been in proximity to an individual resulted positive for the pathogen causing the infectious disease, and requesting these other individuals to follow a quarantine period.

For example, bluetooth based mechanisms, contact tracing systems or similar can be used to individuate and notify other individuals, that they have been in close proximity to an infected individual, and thus have preferably to be subjected to the analysis. Through an appropriate application, the smartphone can identify the infection status of the sample, provide information on necessary isolation measures to the infected individual, and trigger the contact tracing applications on other smartphones, which have recently been in close proximity to the infected person, in order to induce these individuals to follow a period of quarantine.

INFECTION FOLLOW-UP

In some embodiments, individuals resulted positive for the pathogen repeat the analysis to detect the pathogen each one - four weeks in the following weeks, or also also after a period comprised between one day to 3 weeks, preferably between one day to 1 week, in order to follow the development of pathogen infection. Preferably, the infected individual receives the test tube with a unique identification label, where he put his body sample, and he sends back the sample tube to the collection center.

Thus, a particular embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes,

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, q) repeating steps f) - p) after a period comprised between one day to 3 weeks, preferably between one day to 1 week. Optionally, in the step h^') the processing further comprises directly determing the concentration of the specfic amplicaton produts, thus h^') may read as follows: h¹) shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products.

A more particular embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, o) requesting an individual resulted positive for the pathogen causing the infectious disease to follow an isolation period and to follow the recommendations for the treatment of the infectious disease; p) notifying other individuals that they have been in proximity to an individual resulted positive for the pathogen causing the infectious disease, and requesting these other individuals to follow a quarantine period; q) repeating steps f) - p) after a period comprised between one day to 3 weeks, preferably between one day to 1 week.

In some embodiments, repeating step f) - p) is after a period comprised between one to twelve weeks, one to four weeks, between one to three weeks, preferably between one to two weeks, preferably between one day to two weeks, preferably between one day to 3 weeks, preferably between one day to 1 week.

In further embodiments, the infected individuals receive one set of sample tubes that can deliver the total sequencing of the virus genome and information about genes in individuals, that are relevant for the immune response, or the total sequencing of the genome or exome of the infected individual.

PATHOGEN

As used herein the term "pathogen" refers to an organism, including a microorganism, which causes an infectious disease in another organism (e.g., animals and plants) by directly infecting the other organism, or by producing agents that causes disease in another organism (e.g., bacteria that produce pathogenic toxins and the like). As used herein, pathogens include, but are not limited to bacteria, protozoa, fungi, nematodes, viroids and viruses, or any combination thereof, wherein each pathogen is capable, either by itself or in concert with another pathogen, of eliciting disease in vertebrates including but not limited to mammals, and including but not limited to humans. As used herein, the term "pathogen" also encompasses microorganisms which may not ordinarily be pathogenic in a non-immunocompromised host. Specific nonlimiting examples of viral pathogens include coronavirus 229E, SARS-CoV-1, MERS-CoV, SARS-CoV- 2, HSV1, HSV2, EBV, CMV, HHV 6, HHV7, HHV8, VZV, hepatitis C, hepatitis B, adenovirus, EEEV, WNE, JCV, BKV, Influenza A (InfA), Influenza B (InfB), lnfA/H5N1 , RSVA, RSVB, Parainfluenza virus (PIV) PIV-1, PIV-2, PIV-3, PIV-4, Rhinovirus, Human Coronavirus OC43 (Cor-OC43), Human Coronavirus NL63 (Cor-NL63), Human metapneumovirus (HMPV), Encephalomyocarditis-Virus EMC (IAC) As used herein, the term "microorganism" includes prokaryotic and eukaryotic microbial species from the Domains of Archaea, Bacteria and Eucarya, the latter including yeast and filamentous fungi, protozoa, algae, or higher Protista. The terms "microbial cells" and "microbes" are used interchangeably with the term microorganism

In the present invention, the pathogen is selected from the group consisting of: a viral pathogen, a bacterial pathogen, a fungal pathogen, a protozoic pathogen or a multicellular pathogen.

Through the parallel use of appropriate primers it is possible to identify many different virus in only one test, and, through the sequencing, to classify the virus for its biologically and medically relevant characteristics.

In some embodiments, the following virus are analysed in parallel in the same reaction / tube: InfA, InfB, lnfA/H5N1 , RSVA, RSVB, PIV-1 , PIV-2, PIV-3, PIV-4, Rhinovirus, Cor-229E, Cor-OC43, Cor-NL63, hMPV and Adenovirus.

A preferred embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes,

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein the pathogen is a viral pathogen, a bacterial pathogen, a fungal pathogen, a protozoic pathogen or a multicellular pathogen.

A more preferred embodiment of the present invention is directed to a method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery, d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by nucleic acid sequencing, or direct end-point determination, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, i) pooling of a plurality of processed samples collected in the test tubes;

L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1 ) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label, wherein the pathogen is coronavirus 229E, SARS-CoV-1 , MERS-CoV, or SARS- CoV-2.

SYSTEM FOR FIGHTING AN INFECTIOUS DISEASE

The present invention further relates to a system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are collected from the individuals personally, by mail, or by delivery, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label.

Moreover, the present invention relates to a system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are provided to the individuals personally, by mail, or by delivery, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, or direct end-point determination, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label, wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label.

The present invention also relates to a system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are provided to the individuals personally, by mail, or by delivery, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, or direct end-point determination, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label, wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence.

Moreover, the present invention relates to a system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are collected from the individuals personally, by mail, or by delivery, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, or direct end-point determination, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label; wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

Moreover, the present invention relates to a system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are provided to the individuals personally, by mail, or by delivery, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label, wherein the test tube further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response.

SYNTHESIS OF MOLECULAR LABELS

The present invention is also directed to a method to synthetise a molecular label identifying a test tube, comprising: a carrier, (e.g a bead) a nucleic acid sequence linker, a nucleic acid sequence attached to said carrier, wherein said nucleic acid sequence uniquely identifies said carrier, optionally a random nucleic acid sequence differing between different molecular labels attached to the same carrier, an adaptor nucleic acid sequence, wherein the molecular label is optionally linked to one or more of the following elements: at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen, at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, wherein the test tube is provided to collect a body sample to analyse to detect the presence of a pathogen by nucleic acid sequence.

More in particular, the present invention is directed to a method to synthetise a molecular label identifying a test tube, comprising: a bead, a nucleic acid sequence linker, a nucleic acid sequence attached to said bead, wherein said nucleic acid sequence uniquely identifyes said bead, optionally a random nucleic acid sequence differing between different molecular labels attached to the same bead, an adaptor nucleic acid sequence, wherein the molecular label is optionally linked to one or more of the following elements: at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen, at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, wherein the test tube is provided to collect a body sample to analyse to detect the presence of a pathogen by nucleic acid sequence.

Also more in particular, the present invention is directed to a method to synthetise a molecular label identifying a test tube, comprising: a bead, a nucleic acid sequence linker, a nucleic acid sequence attached to said bead, wherein said nucleic acid sequence uniquely identifyes said bead, optionally a random nucleic acid sequence differing between different molecular labels attached to the same bead, an adaptor nucleic acid sequence, wherein the molecular label is optionally linked to one or more of the following elements by an enzymatic procedure: at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen, at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, wherein the test tube is provided to collect a body sample to analyse to detect the presence of a pathogen by nucleic acid sequence, wherein the enzymatic procedure is selected from the group comprising ligation, and site specific recombinases.

Still more in particular, the present invention is directed to a method to synthetise a molecular label identifying a test tube, comprising: a bead, a nucleic acid sequence linker, a nucleic acid sequence attached to said bead, wherein said nucleic acid sequence uniquely identifyes said bead, optionally a random nucleic acid sequence differing between different molecular labels attached to the same bead, an adaptor nucleic acid sequence, wherein the molecular label is optionally linked to one or more of the following elements by an enzymatic procedure: at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen, at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, wherein the test tube is provided to collect a body sample to analyse to detect the presence of a pathogen by nucleic acid sequence, wherein the enzymatic procedure is selected from the group comprising ligation, and site specific recombinases.

A molecular label identifying a test tube can, for instance, be produced by constructing beads carrying DNA sequences consisting of the following elements (5’ to 3’): a common sequence to be used later to introduce sequence elements required for the sequencing process (library arms) by deterministic synthesis, a sequence identifying each bead by combinatorial synthesis (in each step, all beads are randomly distributed to reaction vessels extending the growing chain by one or more bases), potentially a random sequence differing between different molecules (by synthesis with a mixture of all 4 bases in each step), an adaptor sequence, used later to introduce different sequence elements to carry out specific functions linked to the unique identifier for the bead.

To be able to introduce multiple functions into the sequences of the bead, enzymatic techniques (ligation, site specific recombinases etc.) can be used. At least two functional elements are introduced: one or more sequences needed for the specific test or tests to be carried out (e.g. Corona virus genome, Influenza genome, host alleles etc.) one or more sequences required to ultimately provide a (known) identifier of the sequence (typically provided through one or more oligonucleotides, which are enzymatically copied to extend the bead oligonucleotide, with the combination of sequences uniquely identifying the sample tube). Description of Figures

Figure 1 : shows schematic procedures of the inventive method for fighting a method for an infectious disease.

Figure 2: shows a sample collection kit containing 1. sterile gargling liguid;

2. intermediate liquid collection cylinder; 3. pipette; 4. double-barcoded sample tube. The collected double-barcorded sample tubes are located in a double-barcoded 96-well rack (5).

EXAMPLES

Example 1

Individuals, who want to be tested for a pathogen like SARS-CoV2, receive a test tube with an identification label such as a barcode, scan the barcode into their smart phone, and collect samples either by a nasal swab or gargle approach into the test tube containing appropriate preservation materials.

The tubes are collected at regular intervals, the pathogen specific (and for the sequencing sppraich also sample specific) oligonucleotides extended (or linked by other enzymatic procedures to the pathogen sequences) in the case of pathogen positive samples, and then either amplified directly based e.g. on an end-point RT- PCR approach, or pooled and sequenced in high throughput sequencing experiments.

The identification labels (e.g. barcodes) characterising the individual body samples resulted positive for the pathogen are then made available through a special web site allowing individuals to get the results of the test. Alternatively, the individuals can be called or notified through other mechanisms.

Example 2

For population wide screens, all individuals resident in a given area to be analysed will receive test kits by mail. Outside of population wide screens, individuals, who want to be tested for a pathogen, visit a location such as their doctor or a pharmacy, and receive appropriate kits with an identification label such as a barcode.

As all individuals are to be tested, all of them receive packages containing multiple test kits to allow e.g. 5 -28 tests, one to seven per week by mail or at distribution centres. In one implementation each kit contains one tube and a swab device. At the time of the test, the individual scans the barcode on the kit package or on the tube, optionally takes a photo of the tube with the barcode and a personal identification document (e.g. identity card or passport) to be able tolater receive a certificate stating that he/she has been tested positive or negative, inserts the swab device into the nose on both sides, rotates it a few (e.g. five) times, inserts it into the tube, washes off the virus by turning the swab a few times, closes the tub, discards the swab, decontaminates the surface of the tube and drops off the tube at a collection center in an SBS carrier.

Filled carriers are then transported to the processing center, to be analysed by end-point RT-PCR or an equivalent amplification-end-point detection method, or, after adding a unique combination of oligonucleotides incorporated into sample specific sequences on DNA molecules, the sequence of which identifies both the result of the test and a sample specific identifier, pooled and sequenced- Tubes there are i

Example 3: Synthesis of molecular labels

To obviate the need for synthesising millions or billions unique long nucleotides, and to increase the range of tests, which can be carried out in the same sample in parallel, we describe here a modified strategy essentially replacing the one molecule carrying all the information by a sequence library based on three identifiers: the barcode on the tube the unique (but unknown) sequence of the bead oligo ("random nucleic acid sequence stretch". the known identifier of the tube, "molecular label" established by copying information from a combination of oligonucleotides, selected in such a way to uniquely define the tube

A molecular label identifying a test tube can by produced by constructing beads carrying DNA sequences consisting of the following elements (5’ to 3’): a common sequence to be used later to introduce sequence elements required for the sequencing process (library arms), by deterministic synthesis, a sequence identifying each bead by combinatorial synthesis (in each step, all beads are randomly distributed to reaction vessels extending the growing chain by one or more bases) optionally a random sequence differing between different molecules (by synthesis with a mixture of all 4 bases in each step) an adaptor sequence, used later to introduce different sequence elements to carry out specific functions linked to the unique identifier for the bead.

To be able to introduce multiple functions into the sequences of bead, enzymatic techniques (ligation, site specific recombinases etc can be used). At least two functional elements are introduced: one or more sequences needed for the specific test or tests to be carried out (e.g. Corona virus genome, Influenza genome, host alleles etc) one or more sequences required to ultimately provide a (known) identifier of the sequence ("random nucleic acid sequence stretch").

Example 4: e-PCR (endpoint PCR) Test

Self-collection procedures by using Alacris PCR Gargle sample collection Kit. Alacris coronavirus PCR Gargle test

The sample collection kit consists of sterile gargling liquid (1), intermediate liquid collection cylinder (2), pipette (3), and double-barcoded sample tube (4) as shown in Figure 2:

The test person gargles with 10 ml of sterile water that is collected in a liquid collection cylinder. Approximately 300 pi of this is transferred into a barcoded sample tube using a disposable pipette.

Laboratory testing

Collecting all samples and preparation of assembly of all samples for ePCR test. All samples are collected in double-barcoded 96-well racks (5) holding 92 samples as shown in Figure 2.

Alacris e PCR Test

Information of primers and proves of Alacis e-PCR Test kit Thermocvcling conditions The 384 plate will be sealed with the PCR plate sealer and placed intothethermoblockwith the following instructions.

The microtiter plate will be removed and scanned to identify the samples before placing into the CLARIOstar Plus Plate reader for data acquisition.

Read-out of the samples The plate readerwill acquire datafrom each individual samplewiththeuseofthefollowing lasers:

The results will be reported as positive, negative or undetermined based on threshold baseline.

The PCR reagents, PCR times und primers are from 3CR Bioscience. The preprocessing, sample processing, sample and tube selection and handling, data processing and evaluation of results, reporting of results are devised by Alacris.

For example - (3CR Bioscience) Samuiel Moeses et al, “Endopoint PCR Dection of Sars-CoV-2 RNA”, (https://doi.Org/10.1101/2020.07.21 .20158337)

Example 4-1: Endpoint Detection of SARS-CoV-2 in swab samples

1. Purpose and Scope

This SOP covers the endpoint detection of SARS-CoV-2 in swab samples

2. Principle and summary of the method

The SOP is based on a one step RT-PCR kit developed by 3CR Bioscience. The Multiplex assay is for the qualitative detection of RNA of the novel Coronavirus (SARS-CoV-2) in human respiratory samples. The procedure allows the detection of two target sequences in the nucleocapsid (N) gene region (N1 and N2) and an internal RNase P control. The results can be analyzed with endpoint or real-time detection.

The three, N1 + N2 + RNaseP, are combined into one multiplex assay, which are amplified and detected in one reaction. The kit is specifically formulated for endpoint fluorescent detection as N1 reports with FAM, N2 reports with HEX and RNase P control reports with ATTO 647. The assay can be used for real-time detection as well.

3. Type of samples

4. General protocol consideration

• All components must be stored at -20°C upon arrival. · Avoid repeated freeze / thaw cycles.

• This product should only be handled by trained laboratory personnel.

• Wear suitable PPE when using the product.

• In case of contact with skin or eyes, wash immediately with water 5. Perfomrance characteristics

The detection limit of Endpoint detection is N1, N2 < 0.2. Values > 0.2 are regarded as positive and possible outcomes are summarized:

Where values are above the agreed value of 0.2 (based on analysis of proficiency testing performance and other local testing data) with satisfactory quality control parameters, including internal control performance, the result is considered valid and should be communicated and a report issued as a final result. Tests should be repeated for patient samples obtained as undetermined, out of range or if there are no results. Additionally, the head of laboratory should be informed.

6. Definitions and abbreviations

N1 - 2 - Nucleocapsid protein 1 and 2.

NTC - No template control

7. Reagents, materials and devices

7.1 Reagents

7.2 Consumables

• 0.5ml tubes: FluidX 96-Format, 0.5ml External Thread, Next-Gen Dual- Coded Tube

• Racks with lid for 0.65ml, 223.02 (50 racks)

• Pipette filter tips 2-1 OOpl

• Low volume microtitre, clear round-bottom 384 well plates

• PCR plate seal film for 384 plates

7.3 Equipment

8. Procedure

Receiving and scanning samples

• Samples will be received by Alacris Theranostics in racks containing the 94 patient samples (2 empty slots for controls).

• The racks and containing tubes will be scanned using the FluidX Perception™ HD LF Whole Rack Reader to record each rack with tube- containing sample with a unique barcode. Processing samples

• The tubes in each rack will be decapped with the FluidX IntelliXcap™.

• Afer decapping, a volume of 4 pi of each patient sample will be collected and will be added to a 384-microtitre plate via the Biomek i7 or other automatic pipettor. Each 384-microtitreplate fits 4 x 94 samples and the respective positive and negative controls.

• The microtiter plate will be prefilled with 6 mI ProbeSure™ Multiplex Master Mix using the Multidrop reagent dispenser.

Thermocvcling conditions

• The 384 plate will be sealed with the PCR plate sealer and placed into the thermoblock with the following instructions.

• The microtiter plate will be removed and scanned to identify the samples before placing into the CLARIOstar Plus Plate reader for data acquisition. Read-out of the samples

• The plate reader will acquire data from each individual sample with the use of the following lasers:

· The results will be reported as positive, negative or undetermined based on threshold baseline.

9. Release of results

The head of laboratory is responsible to verify the protocol and to release the results to confirm the correctness and possible further steps by its signature. In case of nonconformities, he or she has to decide about necessary actions. In absence of the laboratory head, his or her deputy is authorized for verification and release. Example 4-2: Endpoint Detection of SARS-CoV-2 in qarqle samples

1. Purpose and Scope

This SOP covers the endpoint detection of SARS-CoV-2 in gargle samples 2. Principle and summary of the method

The three, N1 + N2 + RNaseP, are combined into one multiplex assay, which are amplified and detected in one reaction. The kit is specifically formulated for endpoint fluorescent detection as N1 reports with FAM, N2 reports with HEX and RNase P control reports with ATTO 647. The assay can be used for real-time detection as well. 3. Type of samples

4. General protocol consideration · All components must be stored at -20°C upon arrival.

• Avoid repeated freeze / thaw cycles.

• This product should only be handled by trained laboratory personnel.

• Wear suitable PPE when using the product.

• In case of contact with skin or eyes, wash immediately with water

5. Perfomrance characteristics

Where values are above the agreed value of 0.2 (based on analysis of proficiency testing performance and other local testing data) with satisfactory quality control parameters, including internal control performance, the result is considered valid and should be communicated and a report issued as a final result.

Tests should be repeated for patient samples obtained as undetermined, out of range or if there are no results. Additionally, the head of laboratory should be informed.

6. Definitions and abbreviations

N1 - 2 - Nucleocapsid protein 1 and 2.

NTC - No template control

7. Reagents, materials and devices

7.1 Reagents

7.2 Consumables

• Racks with lid for 0.65ml, 223.02 (50 racks)

• Pipette filter tips 2-1 OOpl

• Low volume microtitre, clear round-bottom 384 well plates

• PCR plate seal film for 384 plates 7.3 Equipment

8. Procedure

Receiving and scanning samples

• The racks and containing tubes will be scanned using the FluidX Perception™ HD LF Whole Rack Reader to record each rack with tube- containing sample with a unique barcode.

• The samples will heat-inactivated at 95°C for 30 min.

Processing samples · The tubes in each rack will be decapped with the FluidX IntelliXcap™.

• Afer decapping, a volume of 4 pi of each patient sample will be collected and will be added to a 384-microtitre plate via the Biomek i7 or other automatic pipettor. Each 384-microtitreplate fits 4 x 94 samples and the respective positive and negative controls. · The microtiter plate will be prefilled with 6 mI ProbeSure™ Multiplex Master

Mix using the Multidrop reagent dispenser.

Thermocvcling conditions · The 384 plate will be sealed with the PCR plate sealer and placed into the thermoblock with the following instructions.

• The microtiter plate will be removed and scanned to identify the samples before placing into the CLARIOstar Plus Plate reader for data acquisition.

Read-out of the samples

• The results will be reported as positive, negative or undetermined based on threshold baseline. 9. Release of results

The head of laboratory is responsible to verify the protocol and to release the results to confirm the correctness and possible further steps by its signature. In case of nonconformities, he or she has to decide about necessary actions. In absence of the laboratory head, his or her deputy is authorized for verification and release.

Claims

1 . A method for fighting an infectious disease caused by a pathogen comprising the following steps: a) providing a plurality of test tubes, each test tube for collecting a body sample of essentially each individual of a population, b) providing one identification label for each test tube, c) providing a plurality of collection centers where the test tubes with the identification labels are provided to the individuals personally, by mail, or by delivery; d) providing a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease, by end-point PCR or equivalent amplification techniques, or nucleic acid sequencing, e) providing at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, f) providing at least one of the test tubes to essentially each individual of a population; g) collecting at least one body sample of the individual in one of the provided test tubes and recording the identification label of said test tube on a communication device of the individual, or through a web based interface, in the database; h) processing the body sample collected in the test tube, wherein processing comprises extension and assembly of nucleic acid sequences, and shipping the processed body sample to one of the analysis laboratories; or h') shipping the test tube to one of the analysis laboratories, and processing the body sample collected in the test tube, wherein processing comprises extension and amplification of nucleic acid sequences, and either directly determining the concentration of the specific amplification products or i) pooling of a plurality of processed samples collected in the test tubes; L) analyzing the pooled processed samples to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing at the analysis laboratory, m) storing the analysis of the processed sample in the at least one database in an anonymized form, n) notifying the analysis of the processed sample to the individual by: n1) notifying the analysis to said individual identified through the identification label by contacting said individual; n2) notifying the analysis to said individual through the communication device of step g) identified through the identification label; n3) granting the individual access to the at least one database through the communication device of step g), wherein the communication device is suitable to connect to the database, and to retrieve the analysis of the body sample of the individual by an identification through the identification label.

2. The method according to claim 1 , wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label, or wherein a molecular label identifying the test tube is assembled at step h) or h') with nucleic acid sequences of the individual, and wherein analyzing the pooled processed samples at step I) further comprises nucleic acid sequencing of said molecular label.

3. The method according to claim 1 or 2, wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify the at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end at least one molecular label comprising at least one nucleic acid sequence.

4. The method according to any one of the claims 1 - 3, wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

5. The method according to any one of the claims 1 - 4, wherein the test tube further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, and wherein analysing at step I) the body sample collected in the test tube further comprises sequencing at least one nucleic acid sequence of the individual relevant for the immune response.

6. The method according to any one of the claims 3 - 5, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen of claim 3 and the at least one reference nucleic acid sequence specific for the pathogen of claim 4, further comprise a random nucleic acid sequence stretch to normalize the analyses.

7. The method according to any one of the claims 1 - 6, further comprising after step n) the following steps o) and p): o) requesting an individual resulted positive for the pathogen causing the infectious disease to follow an isolation period and to follow the recommendations for the treatment of the infectious disease; p) notifying other individuals that they have been in proximity to an individual resulted positive for the pathogen causing the infectious disease, and requesting these other individuals to follow a quarantine period.

8. The method according to any one of the claims 1 - 7, further comprising after step n) or step p) the following step q): q) repeating steps f) - n) or steps f) - p) after a period comprised between one day to 3 weeks, preferably between one day to 1 week.

9. The method according to any one of the claims 1 - 8, wherein the pathogen is a viral pathogen, a bacterial pathogen, a fungal pathogen, a protozoic pathogen or a multicellular pathogen.

10. The method according to any one of the claims 1 - 9, wherein the pathogen is coronavirus 229E, SARS-CoV-1, MERS-CoV, or SARS-CoV-2.

11. A system for fighting an infectious disease caused by a pathogen comprising: a) a plurality of test tubes for collecting a body sample of essentially each individual of a population, b) one identification label for each test tube, c) a plurality of collection centers where the test tubes with the identification label are collected from the individuals personally or by mail, d) a plurality of analysis laboratories for analyzing the body sample collected in the test tube to detect the presence of the pathogen causing the infectious disease by nucleic acid sequencing, e) at least one database for storing in anonymized form the analysis of the body sample of a certain test tube connected to a certain identification label, and f) a plurality of communication devices suitable to record the identification label connected to a certain test tube, or a plurality of communication devices suitable to record the identification label connected to a certain test tube and to receive the analysis of the individual body sample by an identification through the identification label; or a plurality of communication devices suitable to record the identification label connected to a certain test tube, to connect to the database, and to retrieve the analysis of the certain body sample by an identification through the identification label.

12. The system according to claim 11, wherein the test tube comprises at least one molecular label comprising at least one nucleic acid sequence and linked to the identification label.

13. The system according to claim 11 or 12, wherein the test tube further comprises at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen to prime and amplify at least one nucleic acid sequence specific for the pathogen, wherein the at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen optionally comprises at one end a molecular label comprising a nucleic acid sequence, and wherein the test tube optionally further comprises at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response.

14. The system according to any one of the claims 11 - 13, wherein the test tube further comprises at least one reference nucleic acid sequence specific for the pathogen to determine the number of nucleic acid sequences specific for the pathogen present in the body sample.

15. A method to synthetise a molecular label identifying a test tube, comprising: a carrier, a nucleic acid sequence linker, a nucleic acid sequence attached to said carrier, wherein said nucleic acid sequence uniquely identifyes said carrier, optionally a random nucleic acid sequence differing between different molecular labels attached to the same carrier, an adaptor nucleic acid sequence, wherein the molecular label is optionally linked to one or more of the following elements: at least one oligonucleotide complementary to at least one nucleic acid sequence specific for the pathogen, at least one oligonucleotide specific for at least one nucleic acid sequence of the individual relevant for the immune response, wherein the test tube is provided to collect a body sample to analyse to detect the presence of a pathogen by nucleic acid sequence.