CN113637798A - Primer and probe for detecting delta 69/70HV deletion mutation site of S gene of new coronavirus Alpha strain and application of primer and probe - Google Patents
Primer and probe for detecting delta 69/70HV deletion mutation site of S gene of new coronavirus Alpha strain and application of primer and probe Download PDFInfo
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Abstract
The invention discloses a primer and a probe for detecting a deletion mutation site of a delta 69/70HV gene of an Alpha strain S of a new coronavirus and application of the primer and the probe, and belongs to the technical field of biology. The primer sequence and the fluorescent probe sequence of the delta 69/70HV deletion mutation site on the S gene in the Alpha mutant strain are optimized, the optimized sequence has the advantages of high specificity, sensitive signal and the like, whether the new crown strain to be detected has the delta 69/70HV deletion S gene or not can be effectively and quickly detected through the delta 69/70del primer and the probe, the method can be used as one of effective proofs for judging whether the new crown strain to be detected is the Alpha mutant strain or not, and a detection means is provided for judging whether the new crown strain to be detected is the Alpha mutant strain or not. After the S gene amplification primer and the fluorescent probe mixture of the new coronavirus Alpha strain and the original strain are added into the reaction tube at the same time, two reaction tubes are not needed to be arranged for carrying out PCR detection respectively, and the S gene of the original strain or the S gene delta 69/70HV deletion type of the Alpha strain of a sample to be detected can be identified in the same reaction tube.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to a primer and a probe for detecting a deletion mutation site of a novel coronavirus Alpha strain S gene delta 69/70HV and application of the primer and the probe.
Background
The World Health Organization (WHO) reports a new coronavirus variant strain with stronger infectivity at 12/14/2020 and is named as B.1.1.7, and at 31/5/2021, the world health organization newly names the new coronavirus variant strain as Alpha mutant strain. Due to its great transmission advantages, the transmission speed of Alpha mutant strains rapidly exceeds that of the original strains of the novel coronavirus. It was reported that the Alpha mutant strain underwent 23 mutations including 14 nonsynonymous mutations (amino acid change), 6 synonymous mutations (amino acid does not change) and 3 deletion mutations in ORF1ab, N gene, ORF8 gene and S (spike) gene, respectively, compared with the novel coronavirus strain (NCBI: NC-045512), and two of the 3 deletion mutations underwent S gene (deletion. DELTA. 69/70HV and deletion. DELTA. 144Y). The amino acid deletion mutation delta 69-70HV of the Alpha mutant strain in the S protein can be beneficial to the immune response of a virus escape host, and is an effective target for identifying the S defect genotype of the Alpha virus strain.
Because the Alpha mutant strain has a transmitting power far higher than that of the original strain of the new coronavirus, the damage is larger. The timely identification of Alph a mutant strains is of great significance. Whole genome sequencing is a gold standard method for identifying viral strains, however, it is time consuming and costly. According to the invention, a primer, a fluorescent probe and a detection method of a delta 69/70HV deletion mutation site on an S gene in an Alpha mutant strain are designed according to Taqman probe fluorescent quantitative PCR (RT-PCR), so that whether a new crown virus strain to be detected has an A69/70HV deletion S gene or not can be detected quickly by the delta 69/70del primer and the probe, and a related detection means is provided for judging whether the new crown virus strain to be detected is the Alpha mutant strain or not.
Disclosure of Invention
Aiming at the defects and problems in the prior art, the invention aims to provide a primer and a probe for detecting a deletion mutation site of a delta 69/70HV of an S gene of a new coronavirus Alpha strain and application of the primer and the probe. The technical advantage of the invention is that the designed delta 69/70HV deletion mutant site primer and probe sequence have the characteristics of simple operation, accuracy, sensitivity, rapidness, high flux and low cost when being used in a detection method based on fluorescent quantitative PCR, and are easy to popularize on a large scale.
The invention is realized by the following technical scheme:
the invention provides a primer for detecting delta 69/70HV deletion mutation sites on S genes of a new coronavirus Alpha strain, which comprises the following components in part by weight: a specific primer pair of delta 69/70HV deletion mutation sites on the S gene of the Alpha strain of the new coronavirus and a specific primer pair of the S gene of the original strain of the new coronavirus.
Wherein, the sequence of a specific primer pair of the delta 69/70HV deletion mutation site on the S gene of the new coronavirus Alpha strain is as follows:
a forward primer: 5'-CTTGGTTCCATGCTATCTC-3' (SEQ. ID. NO. 1);
reverse primer: 5'-ATTATGTTAGACTTCTCAGTGG-3' (SEQ. ID. NO. 2);
the sequence of the novel coronavirus original strain S gene specific primer pair is as follows:
a forward primer: 5'-CTTGTAATGGTGTTGAAGGTT-3' (SEQ. ID. NO. 3);
reverse primer: 5'-CTGGTGCATGTAGAAGTTCAA-3' (SEQ. ID. NO. 4).
The primer group for detecting the delta 69/70HV deletion mutation site on the S gene of the new coronavirus Alpha strain is designed according to the characteristics of PCR amplification reaction, and the target gene fragment can be efficiently amplified under the rapid condition by adopting the amplification primer pair, so that the primer group has the advantages of rapidness, high sensitivity and strong specificity, and can be widely applied to the fields of hospitals, inspection centers, entry and exit and the like which need to be rapidly screened on site.
The invention also provides a probe for detecting the deletion mutation site of the S gene delta 69/70HV of the new coronavirus Alpha strain, which comprises a specific fluorescent probe of the deletion mutation site of the S gene A69/70HV of the new coronavirus Alpha strain and a specific fluorescent probe of the S gene of the original strain of the new coronavirus.
Wherein, the specific fluorescent probe sequence of the delta 69/70HV deletion mutation site on the S gene of the new coronavirus Alpha strain is as follows: 5'-AACCCTGTCCTACCATTTAATGATG-3' (SEQ. ID. NO. 5);
the sequence of the specific fluorescent probe of the S gene of the original strain of the new coronavirus is as follows: 5'-CAACCCACTAATGGTGTTG GTTACC-3' (SEQ. ID. NO. 6).
The specific fluorescent probe of the delta 69/70HV deletion mutation site on the S gene of the new coronavirus Alpha strain is marked with an F AM fluorescent group at the 5 'end and a TAMRA quenching group at the 3' end;
the specific fluorescent probe of the S gene of the original strain of the new coronavirus is marked with a HEX fluorescent group at the 5 'end and marked with a BHQ1 quenching group at the 3' end.
The invention also provides application of the primer group and the fluorescent probe in fluorescent quantitative PCR analysis of the new coronavirus.
In application, the concentrations of the S gene specific primer pairs of the Alpha strain S gene and the original strain S gene are both 0.4 mu mol/L, and the concentrations of the specific probes of the Alpha strain S gene and the original strain S gene + are both 0.2 mu mol/L. In the detection, specific primers and probes of the S gene of the Alpha strain and the S gene of the original strain are mixed and then added into the same reaction tube.
After the novel coronavirus Alpha strain and the S gene amplification primer and the fluorescent probe mixture of the original strain are added into a reaction tube at the same time, the S gene of the original strain or the S gene A69/70HV deletion type of the Alpha strain of a sample to be detected can be identified in the same reaction tube.
Compared with the prior art, the invention has the beneficial effects that:
(1) according to the invention, a primer sequence and a fluorescent probe sequence of a delta 69/70HV deletion mutation site on an S gene in an Alpha mutant strain are optimally designed according to a Taqman probe method fluorescent quantitative PCR (RT-PCR), the optimally obtained sequence has the advantages of high specificity, sensitive signal and the like, whether a new crown virus strain to be detected has the delta 69/70HV deletion S gene or not can be rapidly detected through a delta 69/70del primer and a probe effectively, the sequence can be used as one of effective proofs for judging whether the new crown virus strain to be detected is the Alpha mutant strain or not, a related detection means is provided for judging whether the new crown virus strain to be detected is the Alpha mutant strain or not, and the defects of long time consumption and high cost in a gold standard method for identifying virus strains in the prior art, namely whole genome sequencing are overcome.
(2) Based on the invention, two reaction tubes are not needed to be arranged for respectively carrying out PCR detection, and the S gene of the original strain or the S gene delta 69/70HV deletion type of the Alpha strain of the sample to be detected can be identified in the same reaction tube.
Drawings
FIG. 1 is a fluorescent quantitative PCR curve of the S gene of the new coronavirus Alpha strain based on a delta 69/70HV deletion mutation site primer and a probe;
FIG. 2 is a fluorescent quantitative PCR curve of S gene of original strain of new coronavirus based on delta 69/70HV deletion mutant site primer and probe.
Detailed Description
The invention will be further described with reference to the accompanying drawings. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Example 1 primers and probes for deletion mutation site of S gene A69/70HV of novel coronavirus Alpha strain
Due to the deletion of amino acids 69 and 70 in the S gene of the novel coronavirus Alpha strain, if primers and probes designed based on the S gene of the original strain of the novel coronavirus are used for carrying out RT-qPCR detection on the Alpha strain, the novel coronavirus Alpha strain can not be detected. Through whole genome sequencing verification, an S-gene target failure (SGTF) is determined as a reliable marker of an Alpha strain. A sequence of the deletion site of the S gene delta 69/70HV of the Alpha strain is selected to design primers and probes, and the primers and the probes can be used as one of effective proofs for judging the Alpha strain.
TABLE 1 primer and probe sequences used in the present invention
| Sequence name | Sequence information (5 '-3') | SEQ.ID.No | |
| Alpha strain S gene forward primer | CTTGGTTCCATGCTATCTC | 1 | |
| Alpha strain S gene reverse primer | ATTATGTTAGACTTCTCAGTGG | 2 | |
| Original strain S gene forward primer | CTTGTAATGGTGTTGAAGGTT | 3 | |
| Original strain S gene | CTGGTGCATGTAGAAGTTCAA | 4 | |
| Alpha strain fluorescent probe | AACCCTGTCCTACCATTTAATGATG | 5 | |
| Original strain | CAACCCACTAATGGTGTTGGTTACC | 6 |
Specifically, an S gene specific probe of an Alpha strain shown in SEQ ID.No.5 is marked with FAM fluorescent group at the 5 'end and marked with TAMRA quenching group at the 3' end; the S gene specific probe of the original strain of the new coronavirus shown in SEQ ID No.6 is marked with HEX fluorescent group at the 5 'end and BHQ1 quenching group at the 3' end.
Example 2 detection of deletion mutation site of S Gene Δ 69/70HV of novel coronavirus Alpha strain
First, the reagent needed for detection
1) Reagent I (1 mL/tube): fluorescent quantitative PCR reaction buffer solution.
2) Reagent II (lyophilized): and simultaneously adding a mixture of Alpha strain and original strain primers and probes (the concentration of the primer pair is 0.4 mu mol/L, and the concentration of the probe is 0.2 mu mol/L).
3) And (3) reagent III: premix EX TaqTM enzyme mixture (TAKARA).
4) Positive quality control product: comprises synthetic novel coronavirus Alpha strain S gene target fragments and novel coronavirus original strain S gene target fragments.
5) Negative quality control product: water treated with diethyl pyrophosphate.
6) Specimen type: nasal swab, pharyngeal swab, alveolar lavage fluid.
Second, nucleic acid extraction
Commercial RNA extraction kits such as silica gel membrane-based spin column method or magnetic bead-based nucleic acid extraction kits are used. And (4) operating according to the instruction, and directly detecting the finally collected RNA or storing the RNA in an ultralow temperature refrigerator at minus 80 ℃ for detection.
Third, amplification reaction
Each tube of complete detection system comprises 12.5 mu L of reagent I, 6 mu L of reagent II, 1 mu L of reagent III, 5 mu L of quality control product or nucleic acid detection specimen to be detected and 0.5 mu L of sterile water.
Fourthly, detection step
1) Reagent preparation
And (3) calculating the total amount of the required reagent I, the required reagent II and the required reagent III according to the total reaction number N (including the number of samples to be detected, 1 part of negative quality control products and 1 part of positive quality control products) required by detection, uniformly mixing, and then packaging into a Bio-Rad qPCR 96 pore plate.
2) Sample application
And respectively adding the sample RNA solution, the negative quality control product and the positive quality control product into the subpackaged PCR reagent, wherein the adding amount is 5 mu L, and uniformly mixing to be detected.
3) Detection on machine
A Bio-Rad fluorescence quantitative PCR instrument is adopted for detection, PrimePCR is selected by Run Type, and a program is set on a protocol sub-interface of a Run Setup interface: reverse transcription at 45 ℃ for 10min, pre-denaturation at 95 ℃ for 10min, denaturation at 95 ℃ for 15s, 60 ℃ for 30s, 40 cycles with Sample Volume set to 25 μ L. Express Load selects QuickPlate _96 wells _ All channels.pltd in the Plate interface, selects Sample adding as a positive quality Control hole site, sets Sample type as Standard, sets Sample type as Negative Control hole site, sets Sample type as Unknown at Sample adding hole site of specimen to be detected, simultaneously selects FAM and HEX in Fluorophores, and sets easily distinguished Color. After the setting is finished, saving the file, and clicking the Start Run under the Start Run main interface to Run.
4) Analysis of results
After the reaction is finished, the system automatically stores the detection data file, checks the curve condition of each sample, and clicks the sample hole to check the corresponding detection result.
Reaction wells with obvious amplification curves are positive results, and reaction wells without obvious amplification curves are negative results.
Example 3 specific detection of primers and probes
RNA fragments containing the full-length S gene of the new coronavirus Alpha strain, the new coronavirus original strain, SARS and MERS coronavirus, respectively, are added into the reaction tube, and the detection is performed according to the steps in example 2, as shown in FIG. 1 and FIG. 2, and the results are shown: amplification curve signals appear in FAM and HEX fluorescence channels in the reaction tubes added with the new coronavirus Alpha strain and the new coronavirus original strain respectively. In the reaction tube added with the RNA fragment of the full-length S gene of SARS coronavirus and MERS coronavirus, both the Ct value of FAM and the Ct value of HEX channel are greater than 30 (shown as negative), which indicates that the designed primer and the fluorescent probe for the Alpha strain and the original strain of the new coronavirus have higher reaction specificity.
Example 4 clinical sample testing
Selecting a nasal swab/throat swab sample which is confirmed to contain a novel coronavirus Alpha strain and an original strain respectively by providing clinical and virus culture methods, carrying out nucleic acid extraction on the sample, carrying out gradient dilution on a sample nucleic acid extracting solution by 10, 100 and 1000 times, carrying out detection by adopting the method in the implementation 2, and simultaneously carrying out negative and positive quality control product detection.
The results show that: 1) in the reaction wells only added with the novel coronavirus Alpha strain, an amplification curve (Ct value is 20) appears in the FAM fluorescence channel, no signal appears in the HEX channel, and the Ct values of the FAM fluorescence channel of the novel coronavirus Alpha strain samples after 10, 100 and 1000-fold gradient dilution show a trend of increasing gradients (Ct values are 22, 26 and 28 respectively), and all show typical positive curves.
2) In the reaction wells only added with the novel coronavirus original strain, an amplification curve (Ct value is 20) appears in the HEX fluorescence channel, no signal appears in the FAM channel, and the Ct values of the HEX fluorescence channel of the novel coronavirus original strain sample after 10, 100 and 1000 times of gradient dilution show a trend of increasing gradient (Ct values are 23, 26 and 29 respectively), and all show typical positive curves.
3) In a reaction hole added with a novel coronavirus original strain and a novel crown Alpha strain, both FAM and HEX fluorescence channels show an amplification curve (Ct values are respectively 20 and 21), and the Ct values of FAM and HEX fluorescence channels of a novel coronavirus original strain sample after 10, 100 and 1000 times of gradient dilution show a gradient increasing trend (Ct values are respectively 22, 25 and 28; )22, 26, 29) exhibit typical positive curves (Ct values greater than 30 are considered negative).
Meanwhile, the negative quality control substance displays a negative signal, and the positive quality control substance presents an obvious amplification curve, which shows that the detection result is effective.
The foregoing merely illustrates preferred embodiments of the invention and is therefore described in some detail and not to be construed as limiting the scope of the invention. It should be noted that, for those skilled in the art, various changes, modifications and substitutions can be made without departing from the spirit of the present invention, and these are all within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Sequence listing
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Claims (4)
1. The primers and the probes for detecting the deletion mutation site of the S gene delta 69/70HV of the Alpha strain of the new coronavirus are characterized in that: the primer comprises a specific primer pair of delta 69/70HV deletion mutation sites on the S gene of the new coronavirus Alpha strain, and the nucleic acid sequence of the specific primer pair is as follows:
a forward primer: 5'-CTTGGTTCCATGCTATCTC-3', as shown in SEQ ID No. 1; and
reverse primer: 5'-ATTATGTTAGACTTCTCAGTGG-3', SEQ ID No. 2;
the probe comprises a specific fluorescent probe of a deletion mutation site of a S gene delta 69/70HV of a new coronavirus Alpha strain, and the nucleic acid sequence of the specific fluorescent probe is as follows: 5 '-FAM-AACCCTGTCCTACCATTTAATGATG-TAMRA-3' as shown in SEQ. ID. NO. 5.
2. The primers and probes according to claim 1, characterized in that: the primers also comprise a new coronavirus original strain S gene specific primer pair, and the nucleic acid sequences of the primer pair are as follows:
a forward primer: 5'-CTTGTAATGGTGTTGAAGGTT-3', as shown in SEQ ID No. 3; and
reverse primer: 5'-CTGGTGCATGTAGAAGTTCAA-3', as shown in SEQ ID No. 4;
the probe also comprises a specific fluorescent probe of the S gene of the original strain of the new coronavirus, and the nucleic acid sequence of the specific fluorescent probe is as follows:
5 '-HEX-CAACCCACTAATGGTGTTGGTTACC-BHQ 1-3' is shown as SEQ ID No. 6.
3. The primers and probes according to claim 2, characterized in that: the specific fluorescent probe shown as SEQ.ID.NO.5 has FAM as the 5 'end labeled fluorophore and TAMRA as the 3' end labeled fluorophore; the 5 'end-labeled fluorophore of the specific fluorescent probe shown in SEQ. ID. NO.6 is HEX, and the 5' end-labeled fluorophore is BHQ 1.
4. Use of the primers and probes according to any of claims 1 to 3, characterized in that: the primer and the probe are used for fluorescent quantitative PCR analysis of the new coronavirus.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113913406A (en) * | 2021-12-14 | 2022-01-11 | 中国人民解放军军事科学院军事医学研究院 | Method for detecting SARS-CoV-269: 70del site |
| CN114015810A (en) * | 2021-12-03 | 2022-02-08 | 浙江大学 | Detection kit for simultaneous detection of coronavirus RNA double genes and identification of mutant strains |
| WO2022194756A3 (en) * | 2021-03-15 | 2022-12-01 | F. Hoffmann-La Roche Ag | Compositions and methods for detecting severe acute respiratory syndrome coronavirus 2 (sars-cov-2) variants having spike protein mutations |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112725537A (en) * | 2020-03-12 | 2021-04-30 | 宁波海尔施基因科技有限公司 | Multiplex real-time fluorescent quantitative PCR (polymerase chain reaction) kit and method for detecting 2019 novel coronavirus and primer probe composition |
| CN113005228A (en) * | 2021-04-06 | 2021-06-22 | 上海君远生物科技有限公司 | Detection kit for synchronously detecting multiple respiratory pathogens and detection method thereof |
| CN113025748A (en) * | 2021-01-14 | 2021-06-25 | 复旦大学附属华山医院北院 | Primer composition and kit for rapidly detecting 69-70del mutation of novel coronavirus |
-
2021
- 2021-08-09 CN CN202110910969.2A patent/CN113637798A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112725537A (en) * | 2020-03-12 | 2021-04-30 | 宁波海尔施基因科技有限公司 | Multiplex real-time fluorescent quantitative PCR (polymerase chain reaction) kit and method for detecting 2019 novel coronavirus and primer probe composition |
| CN113025748A (en) * | 2021-01-14 | 2021-06-25 | 复旦大学附属华山医院北院 | Primer composition and kit for rapidly detecting 69-70del mutation of novel coronavirus |
| CN113005228A (en) * | 2021-04-06 | 2021-06-22 | 上海君远生物科技有限公司 | Detection kit for synchronously detecting multiple respiratory pathogens and detection method thereof |
Non-Patent Citations (1)
| Title |
|---|
| NATALI VEGA-MAGAÑA等: "RT-qPCR Assays for Rapid Detection of the N501Y, 69-70del, K417N, and E484K SARS-CoV-2 Mutations: A Screening Strategy to Identify Variants With Clinical Impact", 《FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022194756A3 (en) * | 2021-03-15 | 2022-12-01 | F. Hoffmann-La Roche Ag | Compositions and methods for detecting severe acute respiratory syndrome coronavirus 2 (sars-cov-2) variants having spike protein mutations |
| CN114015810A (en) * | 2021-12-03 | 2022-02-08 | 浙江大学 | Detection kit for simultaneous detection of coronavirus RNA double genes and identification of mutant strains |
| CN113913406A (en) * | 2021-12-14 | 2022-01-11 | 中国人民解放军军事科学院军事医学研究院 | Method for detecting SARS-CoV-269: 70del site |
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