CN110305971A - Application of the staphylococcus epidermis MLST parting in legal medical expert's microbial identification - Google Patents
Application of the staphylococcus epidermis MLST parting in legal medical expert's microbial identification Download PDFInfo
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Abstract
The invention discloses application of the staphylococcus epidermis MLST parting in legal medical expert's microbial identification.The present invention provides application of the 1) staphylococcus epidermis in the microorganism label as legal medical expert's individual identification；2) application of the staphylococcus epidermis as microorganism label in forensic identification individual；3) staphylococcus epidermis traces the application in individual source in forensic identification as microorganism label；The present invention passes through the molecule parting of MLST parting research human skin staphylococcus epidermis, can can not only prove whether using staphylococcus epidermis as can be with the microorganism label of individual identification, simultaneously because MLST also has region, also the possible region source of staphylococcus epidermis can be disclosed by its molecule parting, trace to the source for legal medical expert microorganism from now on and good methodology deposit is provided.
Description
Technical field
The invention belongs to legal medical expert's field of microbial identification, are related to staphylococcus epidermis MLST parting in legal medical expert's microbial identification
In application.
Background technique
In recent years, huge achievement has been taken to the research of human-body biological material evidence in forensic science research field, wherein DNA
Evidence has become most one of strong evidence in all kinds of case material evidences.But with the raising of criminal counter-investigation ability,
The difficulty that valuable DNA can be extracted in cases is increasing, along with some difficult cases are left at the scene
Biological evidence content and its small, detection success rate is low, forces to develop and finds new technical method and is applied to forensic science
Field.Wherein the research of non-source of people biological evidence has become the hot spot and field in urgent need of strengthening that forensic science is studied instantly.And
Microorganism biological material evidence is increasingly becoming the new of forensic science research because of generality, sensitivity and specificity existing for it in recent years
Hot fields.
It is newest research shows that human skin surface distribution bacterium Quantityanddiversity considerably beyond former people's
Estimation, and high individual difference is distributed in the bacterial community of body surface, like each individual be owned by it is a set of unique
" microorganism fingerprint " (microbial fingerprint).The bacterium of these body surfaces attachment can be in contact process transfer
Object surface is moved on to, and the bacterium by transfer can be saved due to the high-resistance to environmental pressure on contact object
For quite a long time.And on skin of the bacterial community (" microorganism fingerprint ") with a people of body surface attachment microenvironment and
Living habit is closely bound up, and bacterial reproduction power is very powerful in addition, can also restore quickly even if after washing one's hands.That is
There is certain correlation in the microbiologic population on the object surface that the microbiologic population of body surface contacted with the individual.Noah
Et al. prediction, can use foundation of the human skin microorganism as medical jurisprudence individual identification.
The research of legal medical expert microorganism at present has not only been rested on microbial physiology biochemistry phenotypic analysis, but quickly into
Enter " genome era ", the development of these technologies can make people in molecular level even genomic level microorganisms
Diversity and complexity, trace to the source strong technical support be provided for the microbiological identification of legal medical expert.But it is quick for application
The base of test sensitivity handles a case for unit, and the microbial genome credit analysis based on high throughput sequencing technologies necessarily faces magnanimity
Microorganism sequencing and data statistic analysis, workload is huge, and cost is very high.No matter this is in financial resources or on the time in public affairs
The popularization and application set up all are huge obstacles.It is not able to satisfy forensic science for qualification result fast and accurately actual demand.
Pulsed-field gel electrophoresis technology (Multilocus sequence typing, MLST) is a kind of based on nucleic acid sequence
The typing of bacteria method of column measurement measures its sequence, analyzes the change of bacterial strain by the multiple house-keeping gene interior segments of PCR amplification
It is different, to carry out parting.MLST is a kind of classifying method being derived by multilocus enzyme electrophoresis (MLEE), is ground by Maiden etc.
Study carefully design, be initially applied in 1998 natural variation meningitis how chromosphere bacterium (Neisseriameningitides), later
It is widely used in other pathogens, environment bacterium and eucaryote.The principle of MLST parting: by measuring 6-10 house keeper's base
Because of the nucleotide sequence of internal 400-600bp, the sequence in each site assigns an equipotential base according to the time sequencing of its discovery
Because of number, the allele number of each plant of bacterium is composed according to the allele that specified sequence arrangement is exactly it, that is, this strain
The sequence type (sequence type, ST) of bacterium.The each ST value obtained in this way represents one group of individual nucleotide sequence letter
Breath.The otherness MLST analysis method of bacterial strain is can be found that by comparing ST value: MLST technology is directed to house-keeping gene design primer
PCR amplification and sequencing are carried out to it, obtain the allele numerical value in each site of each bacterial strain, then carries out allele map
(allelic profile) or sequence type (sequence types, STs) identification, match further according to allele map use
Clustering is carried out to the methods of difference matrix (matrix pair-wise differences) building dendrogram.MLST points
Analyse conceptual design element: (1) bacterial strain of the selection Jing Guo preliminary screening；(2) selection has the gene loci of specific characteristic；(3) it sets
Meter is used for the primer of gene magnification and sequencing.It, can be direct for the bacterium of existing maturation MLST scheme in practice
From acquisition parting scheme in MLST database (http://pubmlst.org/).
Compared with conventional molecular biological classifying method, MLST has higher resolving power, can be divided into bacterium of the same race more
More hypotypes, and determine Phylogenetic Relationships between different ST types and contacted with disease.MLST is easy to operate, as a result can
The comparison for quickly obtaining and being convenient for different experiments room, has been used to the epidemiological surveillance and Study on Evolution of various bacteria.With
The quickening of sequencing speed and the reduction of cost, and the development of analysis software, MLST is increasingly becoming the conventional parting side of bacterium
Method.MLST is more and more by as the common tool that can be carried out international Strain comparison.MLST has become bacterium at present
A kind of important method of molecule epidemic disease-ology research can be compared by database and the result of study of other countries and regions
It is right, to more comprehensively recognize the feature of this area bacterium prevalence.
Staphylococcus epidermis (Staphylococcus epidermidis) is to be originated in largely depositing on human epidermal skin
A kind of gram-positive bacterium.In the skin of human body, nasal cavity, the positions such as vagina are parasitic, belong to normal flora type.To antibiosis
Element is insensitive.It is most commonly seen at skin lesion or wound.It is also possible to which the activity by human body is transferred in environment and air.
That is, it has the popularity being distributed in very normal population, transferability, these features make it is considered that it may
It can be a good legal medical expert microorganism individual identification label.It mainly is concentrated on for the research of staphylococcus epidermis at present
As hospital a kind of important infectious bacteria and the epidemiological survey that carries out.Whether staphylococcus epidermis has in healthy population
Difference in height between individual is anisotropic, if there are regional differences, whether staphylococcus epidermis can move with the region of same individual
The passage of dynamic and time is morphed, and these problems are to determine that can staphylococcus epidermis really become the individual identification of forensic identification
The key of microorganism label.
Summary of the invention
In order to carry out Individual identification in medical law fields, individual identification microorganism label is found, the present invention provides following technology
Scheme:
It is an object of the present invention to provide following purposes.
The present invention provides following 1) -5) in it is any in application:
1) application of the staphylococcus epidermis in the microorganism label as legal medical expert's individual identification；
2) application of the staphylococcus epidermis as microorganism label in legal medical expert's individual identification；
3) staphylococcus epidermis traces the application in individual source in forensic identification as microorganism label；
4) substance for carrying out MLST parting to staphylococcus epidermis is preparing the application in forensic identification individual products；
5) substance of MLST parting is carried out to staphylococcus epidermis in preparing the individual derived products of forensic identification retrospect
Using.
Above-mentioned staphylococcus epidermis derives from Healthy People body surface.
Another object of the present invention is to provide a kind of method of microorganism label for obtaining legal medical expert's individual identification.
Method provided by the invention, includes the following steps:
MLST parting is carried out to the superficial microbes of multiple test individuals respectively, obtains the superficial microbes of multiple individuals
ST value, if the ST value of the superficial microbes is variant in the multiple test individual, the superficial microbes are or wait
It is selected as the microorganism label of legal medical expert's individual identification.
In the above method, the method that the superficial microbes to multiple test individuals carry out MLST parting respectively includes such as
Lower step:
1) superficial microbes of multiple test individuals are identified in separation；
2) superficial microbes are expanded respectively with the corresponding MLST primer of the house-keeping gene of the superficial microbes, obtain
The amplified production of house-keeping gene；
3) amplified production of the house-keeping gene is sequenced, then determines and manages after submitting MLST database to compare sequencing result
The allele spectrum of family's gene；The superficial microbes of each test individual are determined further according to the allele spectrum of the house-keeping gene
ST value, realize MLST parting.
In the above method, the superficial microbes are one or more；
Or, the house-keeping gene is one or more.
In the embodiment of present aspect, the superficial microbes are the staphylococcus epidermis of body surface；The house-keeping gene is
Following 7 genes: carbamate kinase-encoding gene arcC, benzene Zhuo dehydrogenase-encoding gene aroE, abc transport body coding
Gene gtrE, DNA mismatch repair protein coding gene mutS, pyrimidine operon modulin encoding gene pyrR, triose phosphate
Isomery enzyme coding gene tpiA and acetyl coenzyme A transacetylase encoding gene yqiL.Its corresponding MLST primer such as 1 institute of table
Show.
Microorganism label by legal medical expert's individual identification of above method preparation is also the scope of protection of the invention.
Following application is the scope of protection of the invention:
The method of the microorganism label of above-mentioned acquisition legal medical expert individual identification distinguishes the application in different test individuals in legal medical expert；
The microorganism label that the above method obtains legal medical expert's individual identification distinguishes the application in different test individuals in legal medical expert；
The microorganism label that the above method obtains legal medical expert's individual identification traces the application in individual source in forensic identification；
The microorganism label that the above method obtains legal medical expert's individual identification is distinguished in different test individual products in preparation legal medical expert
Application；
The above method obtains the microorganism label of legal medical expert's individual identification in preparing the individual derived products of forensic identification retrospect
Application.
Above-mentioned application are as follows: MLST parting is carried out to the microorganism label, obtains the microorganism label of different test individuals
ST value, realize and distinguish different test individuals.
Another object of the present invention is to provide a kind of method that legal medical expert distinguishes different test individuals.
Method provided by the invention, includes the following steps:
1) the microorganism label of legal medical expert's individual identification is obtained with the method for the microorganism label of above-mentioned identification individual；
2) MLST parting is carried out to the microorganism label, obtains the ST value of the microorganism label of different test individuals, it is real
Now distinguish different test individuals.
3rd purpose of the invention provides a kind of method that legal medical expert distinguishes different test individuals.
The method that legal medical expert provided by the invention distinguishes different test individuals includes the following steps: to body surface skin to be measured
Staphylococcus carries out MLST parting, obtains the ST value of the staphylococcus epidermis of different test individuals, realize legal medical expert distinguish it is different to
Survey individual.
3rd purpose of the invention is to provide a kind of product identified or distinguish individual identification.
Identification or difference individual products provided by the invention, carry out MLST including the microorganism label to legal medical expert's individual identification
The substance of parting.
The said goods further include the substance for obtaining the microorganism label of legal medical expert's individual identification.
Above-mentioned substance is the MLST that the substance and detection microorganism house-keeping gene of MLST parting are carried out to the microorganism of individual
Primer.
The above-mentioned methods and applications being related to are non-disease diagnoses and treatment purposes.
The present invention relates to the staphylococcic separation identifications of human epidermal, the determination and its amplification of MLST parting gene, really
Determine the specific parting of MLST of its skin surface staphylococcus epidermis of different human body, final certification staphylococcus epidermis can be used as method
One of the label of microorganism individual identification is cured, and MLST classifying method is the identification side suitable for legal medical expert microorganism individual identification
Method.
The present invention is exactly to screen in the normal microbial population of human skin surface universal on the basis of early-stage study
With a large amount of existing representative one or more of bacteriums, in-depth study is carried out with the technology of molecular biology, is sought
Suitable bacteria molecule identification method is found, whether the specified germ parting for verifying its skin surface between different individuals has
Whether body otherness, changing over time has stability.If the specific bacteria for meeting above-mentioned two condition, which is treated as one, to be had
The microorganism label of value.One microorganism label is similar to an allele seat of huamn autosomal.Eventually by several
The permutation and combination of a microorganism label forms a set of microbial molecules classification system that can be used for legal medical expert's individual identification, final real
Individual identification is now carried out by microbial process.This maximum advantage of research method is aiming at single or certain several bacterium
Molecule parting is carried out, as a result reliably, analysis method is easy, mature, can be to obtain a result in the short time.Eliminate many and diverse, sea
The bioinformatic analysis of amount.It is very suitable for the actual demand of forensic science.
The present invention passes through the molecule parting of MLST parting research human skin staphylococcus epidermis, can not only prove whether
It can be using staphylococcus epidermis as can may be used also with the microorganism label of individual identification simultaneously because MLST also has region
The possible region source of staphylococcus epidermis is disclosed by its molecule parting, traces to the source for legal medical expert microorganism from now on and good side is provided
Science of law deposit.The acquisition of a large amount of sequencing results can provide largely simultaneously for the special SNP site of microorganisms label from now on
Data and reference reduce inspection cost and detection automation are made to provide technical support to detect microorganism label with SNP from now on.
Detailed description of the invention
Fig. 1 is as forensic identification microorganism label staphylococcus epidermis MLST parting process step.
Fig. 2 is sample microbial enrichment culture after acquisition.
Fig. 3 is staphylococcus epidermis blood plate isolation and purification culture.
Fig. 4 is that ovobiocin sensitivity screens staphylococcus epidermis.
Fig. 5 is the identification of staphylococcus epidermis 16srRNA.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1, staphylococcus epidermis are as the microorganism tag recognition individual difference in forensic identification
Fig. 1 is the staphylococcus epidermis MLST parting process step as forensic identification microorganism label.
The object of the present invention is achieved like this: carrying out separation mirror firstly for the staphylococcus epidermis of human skin surface
It is fixed, followed by the determination and its amplification of MLST parting gene, sequencing finally determine human sample's table according to the gene order of measurement
The specific parting of skin staphylococcus MLST, it is final to determine that human epidermal staphylococcus can be used as the microorganism label in forensic identification.
One, the MLST parting of staphylococcus epidermis
1, the separation identification of staphylococcus epidermis
1) it, samples
Using cotton swab method, a small amount of sterile saline is dipped to multiple shown in table 1 with the aseptic cotton carrier after sterilizing
Article after suspect's test individual finger pulp loss or individual use wipes, and cotton swab is then put back to mating test tube
In, it marks.Second step experiment should be carried out after acquisition as early as possible, is temporarily saved if subsequent experimental cannot be carried out and should be placed on 4 DEG C.
2) it, cultivates
5mL 7.5% sodium chloride broth fluid nutrient medium (beef extract 5g, sodium chloride is added in above-mentioned 1) acquire in sample cell
75g, tryptose powder 10g, surplus is water), after mixing, 37 DEG C of shaking table, 200rpm is cultivated for 24 hours (Fig. 2) for oscillation.It will cultivate
The streak inoculation of liquid culture dilution method in blood agar plate, 37 DEG C of cultures are for 24 hours (Fig. 3).
By above-mentioned cultured monoclonal colonies, chooses 10-20 monoclonal colonies switching and spread cultivation to 7.5% sodium chloride meat
Soup solid medium, 37 DEG C of cultures are for 24 hours.
3), ovobiocin sensitivity screens staphylococcus epidermis
By the 2) thallus that step obtains be seeded in 7.5% sodium chloride broth solid medium respectively and be added to 0.65 μ g/
On 7.5% sodium chloride broth solid medium of mL ovobiocin, 37 DEG C of cultures are for 24 hours.Wherein do not adding 7.5% chlorine of ovobiocin
Change sodium meat soup cultured on solid medium and containing non-growing bacterium on 7.5% sodium chloride broth solid medium of ovobiocin
It falls as doubtful staphylococcus epidermis (Fig. 4).
4) staphylococcus epidermis is identified using 16sRNA
By 3) the doubtful staphylococcus epidermis of step be transferred to 37 DEG C of 7.5% sodium chloride broth fluid nutrient medium of 20mL,
200rpm is cultivated for 24 hours.Genomic DNA is extracted, as template, PCR amplification 16sRNA (Primer1:
AGAGTTTGATCCTGGCTCAG, Primer2:GGTTACCTTGTTACGACTT), amplified production 0.8% Ago-Gel electricity
Swimming detection (Fig. 5), sequencing result are sequence 1, are determined as staphylococcus epidermis after comparing on genbank.
2, PCR amplification
The gene of staphylococcus epidermis MIST parting is following 7 genes: arcC (coding carbamate kinases, Gene
ID:1056966, updated on 30-Jan-2018), aroE (coding benzene Zhuo acidohydrogenase, Gene ID:1057358,
Updated on 6-Sep-2017), gtrE (coding abc transport body, Gene ID:1056358, updated on 6-Sep-
2017), mutS (coding DNA mismatch repair protein, Gene ID:1057646, updated on 30-Jan-2018), pyrR
(coding pyrimidine operon modulin, Gene ID:1057548, updated on 20-Aug-2016), tpiA (coding phosphoric acid
Triose isomerase, Gene ID:1057780, updated on 30-Jan-2018), yqiL (encoding acetyl coacetylase acetyl transfer
Enzyme, Gene ID:1057154, updated on 6-Sep-2017).
The DNA molecular for extracting the staphylococcus epidermis that 1 obtains, expands above-mentioned 7 genes with following primer respectively.
Table 1 is staphylococcus epidermis MLST house-keeping gene and its corresponding MLST primer
Primer sequence in upper table is the corresponding MLST primer of respective house-keeping gene, be originated from MLST database (http: //
www.mlst.net/)。
Wherein used annealing temperature is respectively as follows: arcC (65 DEG C), aroE (63 DEG C), gtrE when this 7 gene magnifications
(63℃)、mutS(53℃)、pyrR(53℃)、tpiA(60℃)、yqiL(58℃)。
Obtain the amplified production (obtaining purpose band) of 7 genes.
3, the determination of MLST parting
By the amplified production bidirectional sequencing of above-mentioned 7 genes, the sequencing result of 7 genes is obtained.
It is determined after submitting MLST database (http://www.mlst.net/) to compare the sequencing result of 7 genes each
The allele spectrum of gene (1-8 of table 2 is arranged).It is composed according to the allele of 7 genes and determines sample mesocuticle staphylococcus
ST type (the 9th column of table 2).
Table 2 is the allele spectrum and the determination of ST parting of staphylococcus epidermidis gene
As can be seen that the staphylococcus epidermis MIST genotyping result of 8 people to be measured is entirely different, and therefore, epidermis grape ball
Bacterium MLST parting can be used for identifying or distinguishing different people to be measured.
Staphylococcus epidermis can be used as the microorganism label of legal medical expert's individual identification.
The ST value of staphylococcus epidermis MLST genotyping result can be used for Different Individual in forensic identification and identify.
Two, individual stability study of the staphylococcus epidermis MLST parting in forensic identification microorganism label
1, detection of the staphylococcus epidermis MLST parting to living between the Different Individual in the same space
Using one method, the same space (the same dormitory of certain colleges and universities big three in table 2 is detected；Community life 3 years) different
Body staphylococcus epidermis MLST parting.
The results are shown in Table 3,
Table 3 is the equipotential base of the same space (the same dormitory of certain colleges and universities big three) Different Individual staphylococcus epidermis MLST parting
Because of spectrum and ST parting
| Name | arcC | aroE | gtrE | mutS | pyrR | tpiA | yqiL | ST |
| Ren | 1 | 5 | 2 | 6 | 2 | 1 | 6 | 73 |
| Sun | 12 | 1 | 9 | 5 | 11 | 4 | 21 | 751 |
| Mr. Wang | 30 | 1 | 60 | 5 | 54 | 5 | 8 | 826 |
| Slowly so-and-so | 2 | 1 | 1 | 1 | 2 | 1 | 1 | 59 |
| Poplar | 2 | 1 | 3 | 2 | 4 | 1 | 7 | 307 |
| Army | 1 | 1 | 2 | 6 | 2 | 42 | 33 | 540 |
It can be seen from the results above that be independent of each other between the Different Individual lived in the same space for a long time, each self-insurance
The ST type for holding oneself is constant.
2, monitoring of the staphylococcus epidermis MIST parting to the different space-times of same individual
Using one method, same individual difference space-time staphylococcus epidermis MLST parting in table 4 is detected.
The results are shown in Table 4, it can be seen that same individual difference space-time staphylococcus epidermis MLST parting is constant.
Table 4 is the MLST parting research of same sample different time staphylococcus epidermis
It can determine that staphylococcus epidermis can be used as forensic identification microorganism label based on these results of study.
Therefore, it from a kind of determination method of the above-mentioned microorganism label that can obtain legal medical expert's individual identification, including walks as follows
It is rapid:
1) microorganism of multiple test individual body surfaces is identified in separation,
2) the corresponding house-keeping gene of microorganism described in MLST primer amplification, obtains the amplified production of corresponding gene；
3) amplified production of the corresponding gene is sequenced, then is determined after submitting MLST database to compare sequencing result every
The allele of a corresponding gene is composed；The ST of microorganism described in determining sample is composed further according to the allele of the corresponding gene
Value, if the ST value of the corresponding microorganism of multiple test individuals is different, which is the microorganism of legal medical expert's individual identification
Label.
Identification Different Individual can be gone with the microorganism label of legal medical expert's individual identification, can also use legal medical expert's individual identification
Microorganism label go to trace individual region source, specifically gone to distinguish different with the ST value of the MLST parting of the microorganism label
Body, or be compared with different geographical individual.
SEQUENCE LISTING
<110>Baoding municipal public security bureau, Chinese Petroleum Univ., Material Evidence Identification Center, Ministry of Public Security
<120>application of the staphylococcus epidermis MLST parting in legal medical expert's microbial identification
<160> 15
<170> PatentIn version 3.5
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<211> 24
<212> DNA
<213> Artificial sequence
<400> 10
gttactaata cttttgctgt gttt 24
<210> 11
<211> 26
<212> DNA
<213> Artificial sequence
<400> 11
gtagaatgta aagagactaa aatgaa 26
<210> 12
<211> 23
<212> DNA
<213> Artificial sequence
<400> 12
atccaattag acgctttagt aac 23
<210> 13
<211> 20
<212> DNA
<213> Artificial sequence
<400> 13
ttaatgatgc gccacctaca 20
<210> 14
<211> 21
<212> DNA
<213> Artificial sequence
<400> 14
cacgcatagt attagctgaa g 21
<210> 15
<211> 25
<212> DNA
<213> Artificial sequence
<400> 15
ctaatgcctt catcttgaga aataa 25
Claims (10)
1. following 1) -5) any application in:
1) application of the staphylococcus epidermis in the microorganism label as legal medical expert's individual identification；
2) application of the staphylococcus epidermis as microorganism label in legal medical expert's individual identification；
3) staphylococcus epidermis traces the application in individual source in forensic identification as microorganism label；
4) substance for carrying out MLST parting to staphylococcus epidermis is preparing the application in forensic identification individual products；
5) substance that MLST parting is carried out to staphylococcus epidermis is preparing the application in the individual derived products of forensic identification retrospect.
2. a kind of method for the microorganism label for obtaining legal medical expert's individual identification, includes the following steps:
MLST parting is carried out to the superficial microbes of multiple test individuals respectively, obtains the ST of the superficial microbes of multiple individuals
Value, if the ST value of the superficial microbes is variant in the multiple test individual, the superficial microbes are or candidate
For the microorganism label of legal medical expert's individual identification.
3. according to the method described in claim 2, it is characterized by: the superficial microbes to multiple test individuals respectively into
The method of row MLST parting includes the following steps:
1) superficial microbes of multiple test individuals are identified in separation；
2) superficial microbes are expanded respectively with the corresponding MLST primer of the house-keeping gene of the superficial microbes, obtain house keeper
The amplified production of gene；
3) amplified production of the house-keeping gene is sequenced, then determines house keeper's base after submitting MLST database to compare sequencing result
The allele of cause is composed；Further according to the ST of the superficial microbes of the determining each test individual of allele spectrum of the house-keeping gene
Value realizes MLST parting.
4. according to the method in claim 2 or 3, it is characterised in that: the superficial microbes are one or more；
Or, the house-keeping gene is one or more.
5. the method according to claim 3 or 4, it is characterised in that: the superficial microbes are the epidermis grape ball of body surface
Bacterium；
The house-keeping gene is following 7 genes: carbamate kinase-encoding gene arcC, benzene Zhuo dehydrogenase-encoding gene
AroE, abc transport body encoding gene gtrE, DNA mismatch repair protein coding gene mutS, pyrimidine operon modulin coding
Gene pyrR, triosephosphate isomerase gene of schistosoma tpiA and acetyl coenzyme A transacetylase encoding gene yqiL.
6. by the microorganism label of the claim 2-5 legal medical expert's individual identification prepared.
7. any the method distinguishes the application in different test individuals in legal medical expert in claim 2-5；
Or, microorganism label as claimed in claim 6 distinguishes the application in different test individuals in legal medical expert；
Or, microorganism label as claimed in claim 6 traces the application in individual source in forensic identification；
Or, microorganism label as claimed in claim 6 distinguishes the application in different test individual products in preparation legal medical expert；
Or, microorganism label as claimed in claim 6 is preparing the application in the individual derived products of forensic identification retrospect.
8. a kind of method that legal medical expert distinguishes different test individuals, includes the following steps:
1) the microorganism label of legal medical expert's individual identification is obtained with the method any in claim 2-5；
2) MLST parting is carried out to the microorganism label, obtains the ST value of the microorganism label of different test individuals, realize area
Not different test individuals.
9. a kind of method that legal medical expert distinguishes different test individuals, include the following steps: to test individual body staphylococcus epidermis into
Row MLST parting obtains the ST value of the staphylococcus epidermis of different test individuals, realizes that legal medical expert distinguishes different test individuals.
10. the product of a kind of identification or difference individual identification, including MLST points are carried out to microorganism label as claimed in claim 6
The substance of type.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113502354A (en) * | 2021-07-14 | 2021-10-15 | 中国医学科学院输血研究所 | Pathogen detection primer and probe set for transplanted patient infection, kit and application |
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| WO2012117431A1 (en) * | 2011-03-03 | 2012-09-07 | Molecular Digital Diagnostics S.R.L. | Method and reagent kit for the identification of biological fluids in a sample |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113502354A (en) * | 2021-07-14 | 2021-10-15 | 中国医学科学院输血研究所 | Pathogen detection primer and probe set for transplanted patient infection, kit and application |
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