WO2022013895A1

WO2022013895A1 - Method to collect and analyze biological samples and corresponding kit

Info

Publication number: WO2022013895A1
Application number: PCT/IT2021/050170
Authority: WO
Inventors: Paolo Galiano
Original assignee: Alifax S.R.L.
Priority date: 2020-07-16
Filing date: 2021-06-01
Publication date: 2022-01-20

Abstract

The invention concerns a kit and a method to collect and analyze a biological sample, in particular saliva, obtained by means of voluntary expulsion from a human or animal organism. The invention can advantageously be used by a user who needs to know in a simple and rapid way the presence of antigens or antibodies of a specific virus causing a particular infection, in particular of the respiratory tract.

Description

METHOD TO COLLECT AND ANALYZE BIOLOGICAL SAMPLES AND CORRESPONDING KIT

FIELD OF THE INVENTION The present invention concerns a method to collect and analyze human or animal biological samples, in particular salivary, nasal/pharyngeal or sputum, to be analyzed using immunochemical or molecular biology diagnostic techniques.

The invention also concerns a corresponding kit to collect and analyze biological samples. BACKGROUND OF THE INVENTION

In the medical or veterinary field, there is a known need to take biological samples in order to subject them to medical analysis. Usually, the sample is taken from a patient in a home or clinical setting in various ways, depending on the type of sample - liquid, semi-liquid, semi-solid or solid - and put in a test tube. The test tube is closed and delivered to the laboratory where the required analysis is carried out.

In this context, the patent document WO2011/073768 in the name of the present Applicant is known. In the case of an infected patient, the clinical need to treat and monitor the infection requires detecting the type of pathogen agent, for example by searching for the relative antigens and/or relative nucleic acids, and the patient’s possible immune response, by searching for related antibodies.

The detection of the pathogen is performed on the sample collected from the site of infection on which analyses or tests, for example molecular biology tests, are typically carried out to detect the nucleic acids of the pathogen, while the analysis of the immune response is performed by serological tests to identify the presence of IgM, IgG or IgA class antibodies or antigens in the patient’s blood or saliva.

In the case of viral infection of the respiratory tract, it is known to use, as a starting material for molecular biology analyses, a swab or a drop of blood for research tests for antigens or antibodies on blood, serum or saliva or sputum. For the saliva sample, there are methods to collect saliva using a sponge to be inserted into the mouth and collecting the sponge soaked in saliva after it has been in the mouth for a certain time. Viral infections, by way of example, caused by viruses of the Coronaviridae class (SARS-CoV, SARS-CoV-2, etc.) can quickly transform a limited outbreak into a global pandemic.

If the ease of spread is accompanied by a high probability of hospitalization of the infected, the infection can turn into an extremely serious health, economic and social problem that makes it essential to adopt anti-contagion and infection monitoring measures in the population.

On the occasion of the spread of the SARS-CoV-2 pandemic, the procedure followed by the health authorities was to test the population through the use of nasal/pharyngeal swabs and serological tests on saliva or blood samples. The extraction yield of nasal/pharyngeal swabs is around 70%, while for serological ones the threshold of the value has not yet been defined, in relation to the antibody response that each infected individual produces with the development of IgM, IgG or IGA class antibodies.

On the occasion of this pandemic, world health structures had a severe shortage of beds available for patient care and emergency solutions such as field hospitals were prepared.

On the occasion of this pandemic, there was also a limited availability of nasal/pharyngeal swabs and a series of delays in carrying out the tests, and consequently there were many fatalities.

The resulting global lockdown created social and economic problems on production activities.

Monitoring the infection is therefore of particular importance since it allows to identify infected people and isolate them before they contribute to the spread of the pathogen.

There is therefore a need to perfect a method and corresponding kit to collect and analyze biological samples that can overcome at least one of the disadvantages of the state of the art.

The purpose of the present invention is therefore to provide a method and corresponding kit to collect biological samples, preferably saliva, which allows a biological sample to be easily collected.

Another purpose is to provide a method and corresponding kit to collect biological samples which allows to preserve the biological sample until the moment of analysis.

Yet another purpose is to provide a method and corresponding kit to collect biological samples which allows to obtain information on the biological sample during the safe transport of the sample itself. It is also a purpose of the present invention to provide a method and corresponding kit to collect saliva samples which allows to collect a biological sample without the aid of a second person.

Another purpose of the present invention is to provide a method and corresponding kit to collect saliva samples which allows the test to be performed directly from the sample test tube, to be used as a self-test, that is, as a test that can be carried out autonomously, also outside conventional healthcare structures.

The Applicant has devised, tested and embodied the present invention to overcome the shortcomings of the state of the art and to obtain these and other purposes and advantages. SUMMARY OF THE INVENTION

The present invention is set forth and characterized in the independent claims. The dependent claims describe other characteristics of the present invention or variants to the main inventive idea.

In accordance with the above purposes, there is provided a method to collect and analyze a biological sample, in particular, saliva, nasal/pharyngeal, sputum, bronchoalveolar lavage (BAL).

The invention concerns a kit and a method to collect and analyze a biological sample, in particular saliva, obtained by means of voluntary expulsion from a human or animal organism. According to one aspect of the present invention, the method provides to collect the biological sample as above by introducing it into a collection member, with a truncated cone shape, inside which a collection zone is made which is delimited at one end by a bottom opposite an aperture, for example but not necessarily elliptical in shape, from which to introduce the biological sample. According to another aspect of the present invention, the method provides to analyze the biological sample inside the collection zone, by taking an analysis unit into contact with it in order to detect antibodies or antigens.

According to another aspect of the present invention, the method provides to insert the analysis unit inside the collection zone as above through a slit made on a closing member, for example a cap or lid, which closes the aperture as above.

According to another aspect of the present invention, the method provides to transfer at least part of the biological sample inside a pocket, or chamber, suitably sized and provided inside the collection zone as above, in order to then at least partly insert the analysis unit in the pocket, or chamber, through the slit as above.

In this way, on the one hand, the volume of biological sample subjected to analysis has a certain and determinate value, for example between about lOOpL and about 150 pL, in order to obtain greater analysis effectiveness. On the other hand, this guarantees a sufficient immersion of the analysis unit in the sample to be analyzed.

The transfer of the determinate portion of biological sample inside the pocket, or chamber, can occur simply by inclining the collection member from a vertical position to a horizontal or sufficiently inclined position, with the pocket or chamber at least partly facing downward, and then return the collection member to a vertical position.

The present invention also concerns a kit for collecting and analyzing biological samples which comprises:

- at least one collection member inside which a collection zone is made, with a truncated cone shape, delimited at one end by a bottom opposite an elliptical shaped aperture from which to introduce the biological sample, preferably a salivary biological sample;

- at least one analysis unit, or dipstick, which can be inserted in the at least one collection member and is configured to contact the biological sample by immersion to search for antigens or antibodies.

According to one aspect of the present invention, the kit can also comprise a sampling unit comprising a nasal/pharyngeal collection swab and at an opposite end a cap configured to close the at least one test tube.

According to another aspect of the present invention, the collection unit can comprise at least one dispensing unit connected to a cap comprising an extruded cannula to dispense the sample into a lateral flow, that is, a known immunochromatographic device for serological analysis.

The present invention can advantageously be used by a user who needs to simply and rapidly know the presence of antigens or antibodies of a specific vims causing a particular infection, in particular of the respiratory tract. The invention is made in such a way as to allow the user to perform, autonomously (self-test) and outside conventional or specialized health facilities, the collection of their biological sample and the consequent analysis immediately after collection.

The present invention can therefore be used to verify the presence of an infection or the exposure to a specific virus.

Given its practicality, the present invention finds advantageous application above all in locations with a large influx of people, such as airports and railway stations who, if infected, contribute to the dangerous global spread of the virus.

This solution does not affect the normal practices for access to or use of such locations, which would therefore not be burdened or slowed down, guaranteeing the normal flow of people since each person can provide to autonomously carry out the collection of their own sample and the analysis. This also makes the procedure safe for third parties who do not have to come into close contact with the potentially infected person or his/her sample.

The kit as above supplies a set of tools that allow it to be used in different modes, based on requirements.

Thanks to the elements of the kit as above it is in fact possible to collect the biological sample in different ways:

1) expelling the saliva sample obtained by spitting by the patient inside the container, facilitated by its funnel shape, in order to perform a dipstick or lateral flow type test

2) inserting a dipstick directly into the saliva sample test tube to detect either the antigen or antibody classes present;

3) use the kit for nasal/pharyngeal sampling with the use of a swab for molecular biology examinations and, through the presence of a cap in connection with the swab, carry out the transport securely.

In particular, the biological sample collection kit and the use of dipsticks, for immersion in the saliva sample, is very useful for mass examinations as required by a pandemic. The kit is, in fact, easy to use in hospitals/health centers or in districts assigned for the sampling of biological samples, but also in unconventional structures such as airports, businesses, military headquarters, railway stations, etc., characterized by a large flow of people.

In fact, the kit, according to one of the possible modes of use, allows a user, by means of the collection member associated with the test tube, to dispose a sample of saliva or sputum, directly from the patient’s spit inside the test tube. This allows to easily obtain a sample amount sufficient to proceed with the analyzes using the dipstick.

Once the sample has been collected, it is possible to proceed with the research analysis of the presence of IgM, IgG or IgA class antibodies or antigens, which can be performed directly in the saliva sampling test tube without the aid of dedicated laboratory instruments or machines and without needing specific technical or medical skills.

In this way, a single or mass screening of the self-test type is possible, that is, performed autonomously by the patient and capable of verifying the presence of identifiable antigen or antibodies.

However, the test tubes can also be used on automatic machines.

With the use of the sampling unit, it is possible to perform a nasal/pharyngeal swab sampling with which to subsequently proceed with the extraction of nucleic acids for molecular biology investigations.

The test tube with the biological sample inside it can be suitably closed by means of a cap that can be screwed to the test tube body and transported to a collection and sorting center or directly to an analysis center in order to proceed with the analysis.

Before transport, the biological sample can be mixed with a suitable sample preservative solution, of a known type.

In particular, the kit can be configured to detect anti-SARS-CoV-2 antigens or antibodies. Optionally, the kit can be configured to detect antigen or antibodies relative to other types of influenza viruses.

In other embodiments, the kit can identify, for example, anti-HIV antibodies or HIV antigen and other viruses in general.

According to another aspect of the invention, the kit also comprises at least one dispensing unit comprising a cannula connected to the test tube cap configured to close the test tube, and an opposite free dispensing end configured to dispense a portion of salivary sample into any lateral flow device whatsoever. BRIEF DESCRIPTION OF THE DRAWINGS These and other aspects, characteristics and advantages of the present invention will become apparent from the following description of some embodiments, given as a non-restrictive example with reference to the attached drawings wherein:

- figs. 1A-1B are a perspective view of a sequence of use of the components of the kit for collecting and analyzing a biological sample in accordance with some embodiments described here;

- figs. 2A-2B are a perspective view of an alternative sequence of use of the components of the kit for collecting and analyzing a biological sample in accordance with other embodiments described here;

- fig. 3 is a perspective view of the components of the kit for collecting and analyzing a biological sample in accordance with some embodiments described here;

- fig. 4 is a perspective view of a collection member in accordance with some embodiments described here;

- fig. 5 is a perspective view of a collection member in accordance with other embodiments described here;

- fig. 6 is a section view of a collection member in accordance with some embodiments described here;

- fig. 7 is a perspective view of a thermoelectric heating/cooling device in association with a test tube with a sampling unit inside it, in accordance with some embodiments;

- fig. 8 is a three-dimensional view of a collection member in accordance with some embodiments described here;

- figs. 9, 10 and 11 are a schematic section representation of a sequence of use of the collection member of fig. 8.

To facilitate comprehension, the same reference numbers have been used, where possible, to identify identical common elements in the drawings. It is understood that elements and characteristics of one embodiment can conveniently be incorporated into other embodiments without further clarifications.

DETAILED DESCRIPTION OF SOME EMBODIMENTS We will now refer in detail to the possible embodiments of the invention, of which one or more non-limiting examples are shown in the attached drawings. The phraseology and terminology used here is also for the purposes of providing non-limiting examples.

The invention concerns a kit and a method to collect and analyze a biological sample S, in particular saliva, obtained by means of voluntary expulsion from a human or animal organism. In the latter case, the intervention of a specialized subject, for example a veterinarian, may be provided.

According to one aspect of the invention, the method provides to collect the biological sample S by introducing it into a collection member 14, with a truncated cone shape, inside which a collection zone 31 is made delimited at one end by a bottom 15 opposite an aperture 38, elliptical in shape, from which to introduce the biological sample S (fig. 1 A).

According to another aspect of the invention, the method provides to analyze the biological sample S inside the collection zone 31 , taking an analysis unit 23 into contact with it in order to detect antibodies or antigens (fig. IB).

The method can provide to dispense an analysis reagent supplied with a special dispenser 24 into the collection zone 31 (fig. 8), and to put the biological sample S in contact with it in order to mix them together, for example by stirring the collection member 14 previously closed with a closing member 40, for example a cap.

In some embodiments, the collection zone 31 can contain inside it the analysis reagent in lyophilized or liquid form.

The analysis reagent has a composition suitable to allow the detection of antigens or antibodies by means of the analysis unit 23.

In some embodiments, it is provided to insert the analysis unit 23 inside the collection zone 31 through a slit 41 made in the closing member 40.

It follows that, once the biological sample S has been collected in the collection zone 31, it is possible to carry out its analysis with the analysis unit 23, eliminating the need to open the collection member 14. This solution makes analyzing the biological sample S safer, since its accidental dispersion in both liquid and also aerosol form is limited or eliminated.

Optionally, it is provided to remove a closing element 45 which closes the slit 41. Since the volume of biological sample S in the collection zone 31 can be low and therefore insufficient to allow a correct immersion of the analysis unit 23, even if the analysis reagent has possibly been added to it, it is desirable to provide an appropriate way to guarantee the correct immersion of the analysis unit 23.

In some embodiments, the invention provides to analyze the biological sample S inside a pocket, or chamber, 50 (fig. 11) provided inside the collection zone 31 and provided with an aperture 51 so it can be filled with the biological sample S so as to make the latter available for analysis with the analysis unit 23.

In particular, the pocket, or chamber, 50 allows to contain a volume between about 100pL and about 150qL of biological sample S.

The pocket, or chamber, 50 is shaped so as to concentrate at least part of the biological sample S in a smaller space than that of the collection zone 31, in particular distributing it vertically rather than horizontally, thus promoting the at least partial, but in any case, sufficient, immersion of the analysis unit 23.

To facilitate the insertion of the analysis unit 23 in the pocket, or chamber, 50 the latter is disposed so that its aperture 51 is aligned with the slit 41.

Once the biological sample S has been collected in the collection zone 31 and the aperture 38 is closed with the closing member 40 (fig. 9), at least part of the biological sample S can be transferred from the collection zone 31 to the inside of the pocket, or chamber, 50 through the aperture 51, simply by suitably inclining the collection member 14 from a substantially vertical position to an inclined or even horizontal position (fig. 10), in which the pocket, or chamber, 50 is facing at least partly downward, and subsequently returning it vertically (fig. 11), in order to insert the analysis unit 23 in the pocket, or chamber 50, through the slit 41 to carry out the antigenic and/or antibody analysis.

As a consequence of the positioning sequence of the collection member 14 represented in the sequence of figs. 9-11, at least part of the biological sample S is transferred from the collection zone 31 without the use of any dedicated instrument and without risk of contact with the biological sample S.

The analysis unit 23 can be configured to be at least partly immersed in the biological sample S. The immersion can occur, in particular, up to a predefined level marked by a marking zone 47 present on the analysis unit 23 which marks the maximum immersion zone that allows to obtain a correct serological analysis.

To collect the biological sample S, in particular a salivary sample, it is provided to take the aperture 38, which, thanks to its elliptical section, facilitates the operation, in correspondence with the mouth.

In a variant shown in fig. 2A, the method can provide to deposit the biological sample S in the collection member 14 provided with an aperture 49 on the bottom 15 closed by a partition 52.

With reference to fig. 2B, the method can provide to associate said collection member 14 with a test tube 12 in order to put the collection zone 31, through the aperture 49 of the bottom 15, in communication with an aperture 30 of a test tube 12 in order to deposit the biological sample S in the test tube 12.

In this way, the collection member 14 can facilitate the introduction of the biological sample S in the tube 12.

In accordance with some embodiments, it can be provided to selectively remove the partition 52 which closes the aperture 49 before associating the collection member 14 with the test tube 12.

Optionally, the analysis unit 23 can be inserted in the test tube 12 even with the collection member 14 associated with it (not shown).

As an alternative to using the analysis unit 23 to perform a serological analysis, some embodiments of the method can provide to dispense a portion of biological sample S into a lateral flow device 100.

With reference to fig. 2C, the method provides to associate a dispensing unit 46 with the aperture 38, in order to dispense a portion of the salivary biological sample S into a lateral flow device 100.

By inverting the collection member 14, the biological sample S is allowed to exit from the collection zone 31 through the dispensing unit 46.

Once the biological sample S has been collected the method can provide to close the collection member 14 with the closing member 40.

With the closing member 40 closed, it can be provided to transport the collection member 14 to the analysis laboratory.

The method can provide, subsequently, or as an alternative to the analysis with the analysis unit 23, a step of extracting nucleic acids from the biological sample S. Some embodiments provide that the step of extracting nucleic acids provides to:

- contact the biological sample S with a solution of proteinase K to obtain a solution in which the proteinase K has a concentration between 1 and 50 mg/ml;

- heat the solution between 40°C and 80°C for a time between 5 and 30 minutes;

- heat the solution between 90°C and 100°C for a time between 2 and 20 minutes;

- cool the solution between 0°C and 10°C for a time of at least one minute.

According to some embodiments, the collection of the biological sample S, as an alternative to voluntary expulsion, can be performed by means of a nasal/pharyngeal swab using a sampling unit 16.

The sampling unit 16 allows to collect a biological sample in cases where it is not possible, or advisable, to use the at least one collection member 14 or a small amount of sample is sufficient, for example in the case of molecular biology analysis.

Once the biological sample S has been collected, the sampling unit 16 can be inserted into the test tube 12 to be transported where required or stored.

Some embodiments of the method can provide to put the biological sample S in contact with a preservative liquid before performing an analysis with the analysis unit 23, or to dispense a portion of the biological sample S into the lateral flow device 100 using, or not, the step of extracting nucleic acids.

The biological sample mixed with the preservative liquid allows it to be transported by reducing or preventing the degradation of the content to be analyzed.

In some embodiments, the serological analysis provides to detect anti-SARS- CoV-2 antibodies and/or SARS-CoV-2 antigens.

With reference to fig. 3, the kit 10 can comprise:

- the collection member 14 inside which the collection zone 31 is made, delimited at one end by the bottom 15 opposite the aperture 38 from which to introduce the biological sample S, preferably a salivary biological sample;

- the analysis unit 23, or dipstick, which can be inserted in the collection member 14 and is configured to contact the biological sample S by immersion to search for antigens or antibodies.

The collection member 14 can comprise a collection portion 14a with a tapered shape, in particular truncated conical, or truncated pyramidal with a preferably elliptical or circular section which perimetrically defines the collection zone 31.

The collection portion 14a can have a longitudinal development substantially similar to the major axis of the aperture 49 or of the bottom 15.

In some embodiments, the closing member 40 is connected, by means of a connection 42 with assisted folding, to the external edge of the at least one collection member 14 in correspondence with the aperture 38 (fig. 6).

According to one aspect of the invention, the pocket, or chamber, 50 is associated with the collection portion 14a which defines its rear wall and is further defined by a front wall 53, facing toward the inside of the collection zone 31, and by two lateral walls 57 and 61.

The pocket or chamber 50 has a substantially tapered shape, for example wedge-shaped, with an open base facing toward the aperture 38.

The analysis unit 23 comprises an immunochromatographic analysis device 26 of the lateral flow type configured to perform a serological test to detect antibodies or antigens.

Lateral flow devices exploit the principle of diffusion by capillarity of liquids, allowing them to pass through a strip of a suitable substrate from one end to the other, for example nitrocellulose, in which a series of substances and/or molecules are adsorbed that react emitting a visible signal if they bind to target molecules, for example, antibodies or antigens.

The analysis unit 23 can be configured to display a different color band according to the type of immunoglobulin detected, in order to make the test result easily and immediately understandable.

This solution allows a person to autonomously perform this test even on him/herself; the result is immediate and readable even by a third party without coming into contact with either the person or the potentially infected biological sample.

For example, the analysis unit 23 can find advantageous use in locations with a large influx of people such as airports, railway stations or means of public transport where it is necessary to check seropositivity to a particular pathogen, or the presence of the pathogen itself, in order to prevent the movement of positive people from causing the spread of the pathogen itself. In this way, the people charged with controlling the boarding zones can supply the kit 10 to the people who are boarding, who, autonomously, can certify their state of health quickly and safely.

The analysis device 26 can comprise a strip of nitrocellulose 27.

In one variant, the nitrocellulose strip 27 can be provided with a visible marking strip 47 which identifies a maximum level of immersion in the biological sample S. The colored marking can preferably be chosen from colors that reduce situations of confusion with the possible chromatic signal that results from the reaction.

According to some embodiments, the analysis unit 23 can comprise a stem 28 which comprises an end associated with the analysis device 26 and a free end suitable to be gripped by a user.

In some embodiments, the analysis device 26 can be configured to detect anti- SARS-CoV-2 antibodies and/or SARS-CoV-2 antigens.

In other embodiments, the bottom 15 can be provided with an aperture 49 configured to put the collection zone 31 in communication with the aperture 30 of a test tube 12.

According to some embodiments, the aperture 49 can be sealed by a selectively removable partition 52, for example by tearing, or it can be made so as to be easily pierced, or in any case suitable to free the aperture 49 (fig. 6).

In some embodiments, the partition 52 can be, for example, a sheet of aluminum, or of other metal or of plastic material, associated with non-permanent adhesive means.

In the presence of the partition 52, the at least one collection member 14 is therefore provided with a closed bottom that allows it to be used as a container in which to deposit the biological sample S without needing to use the test tube 12.

In accordance with fig. 4, the bottom 15 is closed and can be flat or have a concavity that facilitates the collection of the biological sample S.

With reference to figs. 5 and 6, the at least one collection member 14 can also comprise a connection portion 32, provided with the aperture 49, which develops from the bottom 15 in the opposite direction to the collection portion 14a. The connection portion 32 is configured to be at least partly inserted in the aperture 30 of the test tube 12.

In some embodiments, the collection portion 14a can be made so that the development of the major axis or of the diameter of the aperture 38 is greater than the longitudinal development.

In one variant, the collection member 14 can comprise a gripping portion 55, for example a rigid tab, protruding from the collection portion 14a and configured to be gripped by the fingers of a user (fig. 1 A).

The tapered shape allows, when in use, to increase the access area to the aperture 30, facilitating the entry of the biological sample into the at least one test tube 12 and therefore the filling operation, preventing dangerous spills or pours, especially in the case of potentially infected material.

The size and shape of the aperture 38 can be substantially coherent with a mouth so that the deposit of the biological sample S is intuitive to prevent smudging on the outside. In particular, the aperture 38 can preferably have an elliptical section.

The closing member 40 can be provided with a dispensing member 46 configured to allow the dispensing of a portion of the biological sample S. The dispensing member 46 can be for example a cannula 33 possibly provided with a top 37 associated by means of a flexible connection element 39.

Alternatively, the top 37 is integrated in the dispensing end of the cannula 33 and is configured to be removed in order to free the dispensing end as above.

In some embodiments, the kit 10 can also comprise at least one test tube 12.

The at least one test tube 12 can be equipped with a cap 13 which can be provided with a collar 43 which comprises an annular rib 44 configured to be associated with an annular groove 36 of the at least one test tube 12.

According to some embodiments, the at least one test tube 12 is made of transparent material to allow, for example, to view the depth of immersion of the analysis unit 23 into the biological sample S.

According to one aspect of the invention, the kit 10 can also comprise at least one sampling unit 16, configured to perform a swab, for example a nasal/pharyngeal swab. The sampling unit 16 comprises a stem 18 which can be inserted in the at least one test tube 12 provided at one end with a collection swab 20 and at an opposite end with a cap 22 (fig. 3). According to some embodiments, the sampling unit 16 can be provided with a cap 22 comprising a clamping collar 48 mating in shape and size with the aperture 30 of the at least one test tube 12.

According to some embodiments, the kit 10 can comprise a thermoelectric heating/cooling device 54 configured to treat the biological sample according to increasing and decreasing temperature ramps, comprising a thermostat unit 56 divided into a heating part 58 and a cooling part 60 (fig. 7).

The heating part 58 and the cooling part 60 are each provided with at least one seating 62 shaped to accommodate the test tube 12. In one variant, each seating 62 can be configured to allow the insertion of the collar 32 of the collection member 14.

Some embodiments can provide that the kit 10 comprises a solution for preserving the biological sample (not shown).

In accordance with some embodiments, the preservative solution consists of a test tube separated from kit 10.

It is clear that modifications and/or additions of parts or steps may be made to the kit 10 and to the method as described heretofore, without departing from the field and scope of the present invention as defined by the claims.

In the following claims, the sole purpose of the references in brackets is to facilitate reading: they must not be considered as restrictive factors with regard to the field of protection claimed in the specific claims.

Claims

1. Method to collect and analyze a biological sample (S) obtained by means of voluntary expulsion from a human or animal organism, characterized in that it collects said biological sample (S) by introducing it into a collection member (14), with a truncated cone shape, inside which a collection zone (31) is made delimited at one end by a bottom (15) opposite an aperture (38), preferably elliptical in shape, from which said biological sample (S) is introduced, and in that it analyzes said biological sample (S) inside said collection zone (31), taking an analysis unit (23) into contact with it in order to detect antibodies or antigens.

2. Method as in claim 1, wherein it is provided to insert said analysis unit (23) inside said collection zone (31) through a slit (41) made on a closing member (40) provided to close said aperture (38).

3. Method as in claim 2, wherein it is provided to transfer at least part of the volume of said biological sample (S) inside a pocket, or chamber, (50) provided inside said collection zone (31) and provided with an aperture (51) to insert, at least partly, said analysis unit (23) into said pocket, or chamber, (50) through said slit (41).

4. Method as in claim 3, wherein it is provided to transfer at least part of said biological sample (S) from said collection zone (31) to said pocket, or chamber, (50) through said aperture (51) by inclining said collection member (14) from a vertical position to a horizontal position, in which said pocket, or chamber, (50) is facing downward, and subsequently returning said collection member (14) to a vertical position.

5. Method as in any claim hereinbefore, wherein, prior to the analysis with said analysis unit (23), it is provided to put said biological sample (S) in contact with an analysis reagent inside said collection zone (31).

6. Method as in any claim hereinbefore, which provides to associate said collection member (14) with a test tube (12) in order to put, through an aperture (49) in said bottom (15), said collection zone (31) in communication with an aperture (30) of a test tube (12) to deposit said biological sample (S) in said test tube (12).

7. Method as in claim 6, which, prior to said association with said test tube (12), provides to remove a partition (52) which closes said aperture (49).

8. Method as in any claim hereinbefore, wherein it is provided to immerse said analysis unit (23) in said biological sample (S) at a predefined level marked by a marking zone (47) present on said analysis unit (23).

9. Method as in any claim hereinbefore, which provides, subsequently or alternatively to said analysis with said analysis unit (23), a step of extracting nucleic acids from said biological sample (S).

10. Method as in claim 9, wherein said step of extracting nucleic acids provides to:

- contact said biological sample (S) with a solution of proteinase K to obtain a solution in which said proteinase K has a concentration between 1 and 50 mg/ml;

- cool the solution between 0°C and 10°C for a time of at least one minute.

11. Method as in any claim hereinbefore, which provides to associate a dispensing unit (46) with said aperture (38), to dispense a portion of said salivary biological sample (S) in a lateral flow device (100).

12. Method as in claim 9, wherein the collection of said biological sample (S) is carried out by means of a nasal/pharyngeal swab using a sampling unit (16).

13. Method as in claim 12, which provides to insert said sampling unit (16) in a test tube (12).

14. Method as in any claim hereinbefore, wherein it is provided to put said biological sample (S) in contact with a preservative liquid before carrying out an analysis with said analysis unit (23) or dispensing a portion of said biological sample (S) in the lateral flow device (100) using, or not, said step of extracting nucleic acids.

15. Method as in any claim hereinbefore, wherein said serological analysis provides to detect anti-SARS-CoV-2 antibodies and/or SARS-CoV-2 antigens.

16. Kit (10) for collecting and analyzing biological samples, characterized in that it comprises: - at least one collection member (14), with a truncated cone shape, inside which a collection zone (31) is made delimited at one end by a bottom (15) opposite an elliptical-shaped aperture (38) from which said biological sample (S) is introduced, preferably a salivary biological sample; - at least one analysis unit (23) which can be inserted in said at least one collection member (14) and configured to contact said biological sample (S) by immersion to search for antigens or antibodies.

17. Kit (10) as in claim 16, wherein the at least one collection member (14) is provided with a closing member (40) configured to selectively close said aperture (38) and provided with a through slit (41) which can be selectively opened and shaped to allow the insertion of said analysis unit (23) in said collection zone (31).

18. Kit (10) as in claim 17, wherein said collection member (14) comprises a pocket, or chamber, (50) provided inside said collection zone (31), having an aperture (51) facing toward said aperture (38) and substantially aligned with said slit (41), wherein said pocket, or chamber, (50) is configured to receive a given volume of said biological sample (S) collected in said collection zone (31) and to partly accommodate said analysis unit (23).

19. Kit (10) as in any claim from 16 to 18, wherein said at least one collection member (14) comprises a collection portion (14a) with a tapered shape, in particular truncated conical, or truncated pyramidal with a preferably elliptical or circular section which perimetrically defines said collection zone (31).

20. Kit (10) as in any claim from 16 to 19, wherein said bottom (15) is provided with an aperture (49) configured to put said collection zone (31) in communication with an aperture (30) of a test tube (12).

21. Kit (10) as in claim 20, wherein a collar (32) develops from said bottom (15) provided with said aperture (49) and configured to be inserted into said aperture (30).

22. Kit (10) as in any previous claim from 16 to 21, wherein said aperture (49) is sealed by a selectively removable partition (52).

23. Kit (10) as in any claim from 16 to 22, wherein said closing member (40) is provided with a dispensing member (46) configured to allow the dispensing of a portion of said biological sample (S).

24. Kit (10) as in any claim from 16 to 23, wherein it also comprises at least one sampling unit (16) comprising a stem (18) which can be inserted into said at least one test tube (12) provided at one end with a collection swab (20) and at an opposite end with a cap (22) configured to close said at least one test tube (12).

25. Kit (10) as in any claim from 16 to 24, wherein said serological analysis unit (23) comprises an immunochromatographic analysis device (26) of the lateral flow type configured to perform a test to detect antibodies or antigens.

26. Kit (10) as in claim 25, wherein said analysis device (26) comprises a nitrocellulose strip (27).

27. Kit (10) as in claim 26, wherein said nitrocellulose strip (27) is provided with a marking zone (47) which identifies a maximum level of immersion of said nitrocellulose strip (27) in said biological sample (S).

28. Kit (10) as in any previous claim from 12 to 27, wherein said analysis device (26) is configured to detect anti-SARS-CoV-2 antibodies or SARS-CoV-2 antigens.

29. Kit (10) as in any previous claim from 12 to 28, wherein it comprises a thermoelectric heating/cooling device (54) configured to treat said biological sample according to increasing and decreasing temperature ramps, comprising a thermostat unit (56) divided into a first part (58) configured to heat and a second part (60) configured to cool.

30. Kit (10) as in claim 29, wherein said heating part (58) and said cooling part (60) are each provided with at least one seating (62) shaped to accommodate said test tube (12) and/or said collar (32) of said collection member (14).