CN109182605A - For detecting the primer and probe, detection method and kit of people's norovirus - Google Patents
For detecting the primer and probe, detection method and kit of people's norovirus Download PDFInfo
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Abstract
The present invention provides the primer and probes for implementing quantitative fluorescence PCR (RT-qPCR) detection people's norovirus, including the first primer group and the first probe for detecting GI group people's norovirus, and the second primer sets and the second probe for detecting GII group people's norovirus.The present invention also provides a kind of methods and kit for detecting people's norovirus, and detection method includes: step 1: extracting sample RNA；Step 2: RT-qPCR detection architecture is set up with the first primer group of GI group people's norovirus and the first probe, with the second primer sets of GII group people's norovirus and the second probe；Step 3: the judgement of testing result is carried out according to fluorescence signal.It reduces the omission factor for the existing people's norovirus to have made a variation and substantially shortens detection cycle, reduce artificial and time cost；Detection reaction volume has also carried out appropriate reduction, reduces detection reagent cost；Testing result differentiates simply, has more practicability.
Description
Technical field
The present invention relates to molecular biology fields, the more particularly, to primer and probe of detection people's norovirus, detection
Method and kit.
Background technique
Norovirus (noroviruses, NoVs) is also known as norwalk virus (Norwalk Virus), class norwalk virus
(Norwalk-like Virus) or small round shape virus (Small Round Structured Virus, SRSV), are under the jurisdiction of embedding
Cup section's norovirus belongs to member, is single strand RNA virus.
According to the difference of viral major structural protein nucleic acid sequence, NoVs is divided into 7 groups (GI-GVII), wherein GI,
GII and GIV group NoVs can infect the mankind, referred to as people's norovirus (human noroviruses, HuNoVs).Epidemic disease
Learn statistics indicate that: HuNoVs (based on GI and GII group) is the main arch-criminal for causing acute nonbacterial gastroenteritis, have morbidity
The features such as rate is high, infective dose is low, strong to extraneous resistance.Disease Control and Prevention Center of the U.S. to the epidemic monitoring of the state the result shows that,
1527 food origin disease events are broken out between 2009-2010 altogether, wherein the pathogen of 790 epidemic situations is HuNoVs, are accounted for total sudden and violent
The 42% of hair event.
It is determined currently, the detection method of HuNoVs is established in virion shape, specific nucleic acid section and major antigen
On the basis of cluster, mainly there are electron microscopy, immunological method and molecular detecting method etc..Chinese patent CN201510487540.1,
CN201510179138.7, CN201310553678.8, CN201310271924.0 etc. contribute to the Molecular Detection of NoVs.
Therefore, it is necessary to research and develop a kind of high sensitivity, accuracy rate is high, easy to operate, time-consuming short detection architecture.
Summary of the invention
In view of the above drawbacks of the prior art, technical problem to be solved by the invention is to provide one kind to be able to detect now
People's norovirus and detection sensitivity it is high, accuracy rate is high, easy to operate and time-consuming short detection primer and probe, detection side
Method and detection kit.It should be noted that the scheme in the present invention does not need to solve all the problems above simultaneously, can only solve
Certainly one or more of problems.
To achieve the above object, one aspect of the present invention provides a kind of primer for detecting people's norovirus and spy
Needle.In a preferred embodiment, for detecting the primer and probe of people's norovirus, including for detecting GI group people promise such as
The first primer group of virus and the first probe and the second primer sets and the second probe for detecting GII group people's norovirus；
Wherein, the first primer group include the first primer as shown in SEQ ID NO:1, the second primer as shown in SEQ ID NO:2 and
The third primer as shown in SEQ ID NO:3；The sequence of first probe is as shown in SEQ ID NO:4；
Second primer sets include the 4th primer as shown in SEQ ID NO:5, the 5th primer as shown in SEQ ID NO:6
With the 6th primer as shown in SEQ ID NO:7；The sequence of second probe is as shown in SEQ ID NO:8.Further, it first visits
5 ' ends of needle are marked with the first fluorescent reporter group HEX, and 3 ' ends are marked with MGB modification group；5 ' ends of the second probe are marked with
Second fluorescent reporter group FAM, 3 ' ends are marked with MGB modification group.
Another aspect of the present invention provides a kind of method for detecting people's norovirus.
In a preferred embodiment, the method for detection people's norovirus includes:
Step 1: sample RNA is extracted；
Step 2: with for detecting GI group people's norovirus the first primer group and the first probe, for detecting GII group people
Second primer sets of norovirus and the second probe establish RT-qPCR detection architecture, carry out RT-qPCR；Wherein, the first primer
Group, the first probe, the second primer sets and the second probe are as described in top；
Step 3: the judgement of testing result is carried out according to fluorescence signal.
Further, the judgment method in step 3 are as follows: have amplified signal before the 40th circulation, then promise containing someone in sample
Such as virus；If the amplification information of different fluorescence signals occurs in amplification, contain different genotype people norovirus in sample.
Further, in step 2, RT-qPCR response parameter are as follows: 42 DEG C of reverse transcriptions 3min, 95 DEG C of denaturation 5s, with laggard
Enter 40 circulations, loop parameter is 95 DEG C of denaturation 3s, and 60 DEG C of annealing extend and fluorescence detection 7s.
Further, in step 2, the reaction system of RT-qPCR are as follows:
Another aspect of the invention provides a kind of method for detecting people's norovirus.
In a preferred embodiment, the kit include for detect the first primer group of GI group people's norovirus and
First probe, the second primer sets and the second probe for detecting GII group people's norovirus；Wherein, the first primer group, first are visited
Needle, the second primer sets and the second probe are as described in top.
Optionally, which further includes following reagent: RT-qPCR Buffer, Ex Taq HS, reverse transcriptase mixture
With RNase-free distilled water.
Further, which detects GI group and GII group people's norovirus using the method for RT-qPCR.
People's norovirus is a kind of RNA virus that variability is strong, if using the inspection of conventional two primers cooperation probe
Survey system, detection coverage rate are lower.Moreover, existing primer and probe is both for pervious viral design, to present
The viral diagnosis effect to have made a variation is poor.
Compared with prior art, the present invention in for the detection primer group of GI or GII genotype all include three primers and
One probe, primer sets include three primers, can reduce the use of degeneracy base, to guarantee preferably to match existing normal
See virus, improves detection coverage rate.The present invention is with following the utility model has the advantages that using primer and probe and detection of the invention
Method detects different genotype people norovirus and reduces needle since primer is greatly improved and having been optimized with probe
Omission factor to the existing people's norovirus to have made a variation simultaneously substantially shortens detection cycle, reduces artificial and time cost；
Detection reaction volume has also carried out appropriate reduction, reduces detection reagent cost；Testing result differentiates simply, with more practical
Property.
It is described further below with reference to technical effect of the attached drawing to design of the invention, specific steps and generation, with
It is fully understood from the purpose of the present invention, feature and effect.
Detailed description of the invention
Fig. 1 is that GI genotype people norovirus RT-qPCR detection method sensitivity evaluation is real in the embodiment of the present invention 3
Test result figure；
Fig. 2 is that GII genotype people norovirus RT-qPCR detection method Evaluation on specificity is real in the embodiment of the present invention 3
Test result figure.
Specific embodiment
Multiple preferred embodiments of the invention are introduced below with reference to Figure of description, keep its technology contents more clear and just
In understanding.The present invention can be emerged from by many various forms of embodiments, and protection scope of the present invention not only limits
The embodiment that Yu Wenzhong is mentioned.
In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition, such as Sambrook etc.
Molecular cloning: described in laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989)
Condition, or according to the normal condition proposed by manufacturer.
The detection method of existing HuNoVs is established to be determined in virion shape, specific nucleic acid section and major antigen
On the basis of cluster, mainly there are electron microscopy, immunological method and molecular detecting method etc., Chinese patent CN201510487540.1,
CN201510179138.7, CN201310553678.8, CN201310271924.0 etc. also contribute to the Molecular Detection of NoVs.
But these detection methods all have the defects that it is as follows: 1) detection needed for primed probe be many years ago viral genetic letter
Breath, but HuNoVs genome easily makes a variation, therefore has detection leakage phenomenon at present；2) detection cycle is long.Therefore, it is necessary to develop a kind of needle
Detection architecture high to present HuNoVs detection sensitivity, accuracy rate is high and easy to operate, time-consuming is short.
Embodiment 1 is used to detect the design of the primer and probe of people's norovirus
Amplimer is designed according to the conserved sequence in different genotype people's norovirus genomic RNA sequence.In GI group
In people's norovirus, selecting accession number GenBank No.KF306212.1 whole genome sequence is standard, and phase is downloaded on NCBI
57 whole genome sequences are closed, conservative gene section therein are found by BLAST, using it as the inspection of GI group people's norovirus
It surveys target gene (the 5288-5427 section of GenBank No.KF306212.1)；In GII group people's norovirus, accession number is selected
GenBank No.KU870455.1 whole genome sequence is standard, and related 132 whole genome sequences are downloaded on NCBI, are led to
It crosses BLAST and finds conservative gene section therein, using it as the detection target gene (GenBank of GII group people's norovirus
The 4970-5100 section of No.KU870455.1).
According to above-mentioned conserved region sequence design primer and probe, determined by experimental result such as table from alternative primer pair
Primer and probe sequence shown in 1:
1 primer and probe sequence of table
In table 1, R represents A or G；Y represents C or T；M represents A or C；H represents A or T or C.
In an alternative embodiment, 5 ' ends of probe LZIP sequence are marked with the first fluorescent reporter group HEX, 3 ' ends
It is marked with MGB modification group, i.e. LZIP:HEX-CGTCCTTAGACGCCATCATCATTTAC-MGB；5 ' end marks of LZIIP sequence
Note has the second fluorescent reporter group FAM, and 3 ' ends are marked with MGB modification group, i.e. LZIIP:FAM-AGCCAGATTGCGATCGCC-
MGB。
The detection method of 2 people's norovirus of embodiment
Primer and probe as described in Example 1 are used in the present embodiment, primer is limited by Jiangsu gold only intelligence biotechnology
Company's synthesis.
The detection method of people's norovirus includes: step 1: the preparation of RNA template；Step 2: using including LZIF, LZIR-
The first primer group, probe LZIP, the second primer sets including LZIIF-A, LZIIF-B, LZIIR and the probe of A and LZIR-B
LZIIP carries out RT-qPCR；Step 3: the judgement of testing result is carried out according to fluorescence signal.
In step 1, sample is diluted 10 times with PBS, 3,000r/min centrifugation 10min take 140 μ L supernatants, are put into
In 1.5mL centrifuge tube；According to HiPure Viral RNA Kit Viral extraction kit operating instruction (Magen, Guangzhou, China)
Nucleic acid extraction is carried out, is finally eluted with the distilled water of 60 μ L RNase-free；After aliquot packing, -80 DEG C are frozen.
Wherein, LZIR-A and LZIR-B is first mixed to form mixed liquor, and mixed liquid concentration is 20 μM；LZIIF-A and LZIIF-B
It is first mixed to form mixed liquor, mixed liquid concentration is 20 μM.
Sequentially add each reaction reagent in above-mentioned reaction system into reaction tube, and using sterile water be template as anti-
The negative control answered.After reaction tube is centrifuged, it is put into BIORAD CFX96qPCR reaction instrument, according to following PCR cycle parameter
Carry out: 42 DEG C of reverse transcriptions 3min, 95 DEG C of denaturation 5s then carry out 40 circulations, and the program of each circulation includes 95 DEG C of denaturation 3s,
60 DEG C of annealing extend 7s (containing fluorescence detection), terminate all operation sequences later.
Judgment method in step 3 are as follows: have amplified signal before the 40th circulation, then norovirus containing someone in sample.By
The fluorescent reporter group of different fluorescence signals is had in probe, if therefore there is the amplification information representative sample of different fluorescence signals
In contain different genotype people norovirus.
The sensitivity evaluation of the detection method of 3 people's norovirus of embodiment is tested
It takes from national disease prevention and control center, Beijing Disease Prevention and Control Centre and Zhejiang Province's disease prevention control
Different genotype people's norovirus diarrhea sample at center processed is 23 parts total, extracts people in the way of step 1 in embodiment 2
Norovirus geneome RNA.After measured, the nucleic acid concentration of geneome RNA is about 0.5*106Copy/ μ L, it is bis- with RNase-free
It steams water and makees 10 times of gradient dilutions, dilute 6 gradients altogether, each dilution gradient takes 2.0 μ L that such as embodiment 2 is added as template respectively
The step of two described in RT-qPCR reaction system, and detected according to the RT-qPCR reaction method in embodiment 2.Its
In, primer and probe is primer and probe shown in embodiment 1.
Testing result is as depicted in figs. 1 and 2.Therefore, determine the detection sensitivity of the detection architecture for 1.0copy/RT-
QPCR reaction system.
The Evaluation on specificity of the detection method of 4 people's norovirus of embodiment is tested
Take people's norovirus and inhuman norovirus sample totally 24 parts (as shown in table 2).
1, it is detected according to the method in embodiment 2
Geneome RNA is extracted in the way of step 1 in embodiment 2.The geneome RNA solution of every part of sample takes 2.0
μ L as template be added to such as the step of embodiment 2 two described in RT-qPCR reaction system, and according in embodiment 2
RT-qPCR reaction method is detected.Wherein, primer and probe is primer and probe shown in embodiment 1.
2, it is detected with commonly using primer and probe at present
Referring to document " Broadly Reactive and Highly Sensitive Assay for Norwalk-Like
Viruses Based on Real-Time Quantitative Reverse Transcription-PCR”(Tsutomu
Kageyama et.al, J Clin Microbiol, 2003,41 (4): 1548-1557) used in primer, probe, reactant
System and response parameter, as the current check experiment for reusing primer and probe and being detected.
The reaction system of RT-qPCR is as follows:
Response parameter is as follows:
42 DEG C of 30min, 95 DEG C of denaturation 5min, subsequently into 40 thermal cycles, loop parameter is 95 DEG C of denaturation 15s, 56 DEG C
Annealing extends 1min.
2 people's norovirus of table and inhuman norovirus sample and testing result comparison
Detected that the results are shown in Table 2 in the way of in embodiment 2.As can be known from Table 2, as long as containing in sample
People's norovirus just has fluorescence signal generation.And people's norovirus RT-qPCR of existing frequently-used primer and probe detects body
It is that in the sample of 17 norovirus containing someone, having 4 can not be detected.Also, it is carried out in the way of in embodiment 2
Detection, Ct value are less than the system of common primer and probe, i.e. its detection sensitivity is higher.It can be seen that the detection side in embodiment 2
Method and the more existing most common people's norovirus RT-qPCR detection architecture of system are compared, and have very high reliability, and advantage is bright
It is aobvious.
The preferred embodiment of the present invention has been described in detail above.It should be appreciated that the ordinary skill of this field is without wound
The property made labour, which according to the present invention can conceive, makes many modifications and variations.Therefore, all technician in the art
Pass through the available technology of logical analysis, reasoning, or a limited experiment on the basis of existing technology under this invention's idea
Scheme, all should be within the scope of protection determined by the claims.
Sequence table
<110>Shanghai Communications University
<120>for detecting the primer and probe, detection method and kit of people's norovirus
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cgtcaytcga cgccatcttc attcac 26
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agccagattg cgatcgcc 18
Claims (9)
1. the primer and probe for detecting people's norovirus, which is characterized in that including for detecting GI group people's norovirus
The first primer group and the first probe and the second primer sets and the second probe for detecting GII group people's norovirus；
Wherein, the first primer group includes the first primer as shown in SEQ ID NO:1, the as shown in SEQ ID NO:2
Two primers and the third primer as shown in SEQ ID NO:3；The sequence of first probe is as shown in SEQ ID NO:4；
Second primer sets include the 4th primer as shown in SEQ ID NO:5, the 5th primer as shown in SEQ ID NO:6
With the 6th primer as shown in SEQ ID NO:7；The sequence of second probe is as shown in SEQ ID NO:8.
2. as described in claim 1 for detecting the primer and probe of people's norovirus, which is characterized in that first probe
5 ' ends be marked with the first fluorescent reporter group HEX, 3 ' ends are marked with MGB modification group；5 ' end labels of second probe
There is the second fluorescent reporter group FAM, 3 ' ends are marked with MGB modification group.
3. a kind of method for detecting people's norovirus, which is characterized in that the described method includes:
Step 1: sample RNA is extracted；
Step 2: with for detecting GI group people's norovirus the first primer group and the first probe, for detecting GII group people promise such as
The second primer sets and the second probe of virus establish RT-qPCR detection architecture, carry out RT-qPCR；Wherein, the first primer group,
One probe, the second primer sets and the second probe are as described in claim 1；
Step 3: the judgement of testing result is carried out according to fluorescence signal.
4. the method for detection people's norovirus as claimed in claim 3, which is characterized in that the judgment method in step 3 are as follows:
There is amplified signal before 40th circulation, then norovirus containing someone in sample；If there is the amplification letter of different fluorescence signals in amplification
Breath then contains different genotype people norovirus in sample.
5. the method for detection people's norovirus as claimed in claim 3, which is characterized in that in step 2, RT-qPCR reaction ginseng
Number are as follows: 42 DEG C of reverse transcriptions 3min, 95 DEG C of denaturation 5s subsequently enter 40 circulations, and loop parameter is 95 DEG C of denaturation 3s, 60 DEG C of annealing
Extension and fluorescence detection 7s.
6. the method for detection people's norovirus as claimed in claim 3, which is characterized in that in step 2, the reaction of RT-qPCR
System are as follows:
7. a kind of detection kit of people's norovirus, which is characterized in that the kit includes for detecting GI group people promise such as
The first primer group and the first probe, the second primer sets and the second probe for detecting GII group people's norovirus of virus；Its
In, the first primer group, the first probe, the second primer sets and the second probe are as described in claim 1.
8. the detection kit of people's norovirus as claimed in claim 7, which is characterized in that the kit further includes following
Reagent: RT-qPCR Buffer, Ex Taq HS, reverse transcriptase mixture and RNase-free distilled water.
9. the detection kit of people's norovirus as claimed in claim 7, which is characterized in that the kit uses RT-
The method detection GI group and GII group people's norovirus of qPCR.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110221080A (en) * | 2019-06-25 | 2019-09-10 | 上海交通大学 | A kind of source of people norovirus immune colloid gold reagent box and cell strain |
| CN110592284A (en) * | 2019-10-12 | 2019-12-20 | 福建谷科生物科技有限公司 | Kit for rapidly detecting norovirus |
| CN113186354A (en) * | 2021-05-28 | 2021-07-30 | 中国科学院微生物研究所 | Kit and special primer for detecting GI.1 type norovirus in clinical sample |
| CN114316009A (en) * | 2020-09-29 | 2022-04-12 | 上海交通大学 | Protein capable of combining multiple viruses and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108085414A (en) * | 2017-12-18 | 2018-05-29 | 北京卓诚惠生生物科技股份有限公司 | Norovirus GI types and GII type detection primer probe groups |
-
2018
- 2018-10-10 CN CN201811176679.4A patent/CN109182605A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108085414A (en) * | 2017-12-18 | 2018-05-29 | 北京卓诚惠生生物科技股份有限公司 | Norovirus GI types and GII type detection primer probe groups |
Non-Patent Citations (2)
| Title |
|---|
| DANLEI LIU等: "Redesigned Duplex RT-qPCR for the Detection of GI and GII Human Noroviruses", 《ENGINEERING》 * |
| PENG TIAN等: "Estimation of Human Norovirus Infectivity from Environmental Water Samples by In Situ Capture RT-qPCR Method", 《FOOD ENVIRON VIROL》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110221080A (en) * | 2019-06-25 | 2019-09-10 | 上海交通大学 | A kind of source of people norovirus immune colloid gold reagent box and cell strain |
| CN110592284A (en) * | 2019-10-12 | 2019-12-20 | 福建谷科生物科技有限公司 | Kit for rapidly detecting norovirus |
| CN114316009A (en) * | 2020-09-29 | 2022-04-12 | 上海交通大学 | Protein capable of combining multiple viruses and application thereof |
| CN114316009B (en) * | 2020-09-29 | 2023-04-25 | 上海交通大学 | Protein capable of combining multiple viruses and application thereof |
| CN113186354A (en) * | 2021-05-28 | 2021-07-30 | 中国科学院微生物研究所 | Kit and special primer for detecting GI.1 type norovirus in clinical sample |
| CN113186354B (en) * | 2021-05-28 | 2023-01-24 | 中国科学院微生物研究所 | Kit and special primer for detecting GI.1 type norovirus in clinical sample |
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