Primer probe combination for detecting six respiratory viruses, kit and application

A primer-probe and kit technology, applied in the field of medical detection, can solve the problems of high heating and cooling process required by testing personnel, time-consuming at least 1.5 hours, frequent and time-consuming experimental operation steps, etc., so as to reduce the number of undiagnosed cases. burden, timely targeted treatment, and the effect of rapid differential diagnosis

Active Publication Date: 2020-04-21
CAPITALBIO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although real-time fluorescent PCR technology has the characteristics of high sensitivity and high specificity, the experiment requires high detection personnel and the experimental operation steps require frequent heating and cooling processes, requiring expensive instruments, and light detection takes at least 1.5 hours above, and most of them

Method used

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  • Primer probe combination for detecting six respiratory viruses, kit and application
  • Primer probe combination for detecting six respiratory viruses, kit and application
  • Primer probe combination for detecting six respiratory viruses, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Embodiment 1, screening and preparation of primers and probe sets

Embodiment 201

[0098] The performance of detection primers and probes determines the pros and cons of the detection effect of the kit. The present invention screens multiple sets of primer-probe combinations for each detection index, and finally screens a set of optimal primer-probe combinations. This example takes the 2019-nCoV detection index as an example to illustrate the primer screening process.

[0099] Primer design:

[0100] Primers and probes were designed for the S gene and N gene of the 2019-nCoV virus (Table 2), and the combination of primers and probes with the best detection effect was screened out through experiments. The primers and probe sequences used were provided by Sangon Bioengineering (Shanghai) Co., Ltd. synthesis.

[0101] Table 2 Primer and probe sequences used to detect 2019-nCoV virus

[0102]

[0103] Continuation

[0104]

[0105] Continuation

[0106]

[0107] Continuation

[0108]

[0109] In the combination of primers and probes mentioned a...

Embodiment 2

[0128] Embodiment 2, kit accuracy and specificity analysis

[0129] Experimental sample:

[0130] In order to verify the accuracy and specificity of the kit, some national reference products of second-generation influenza A / B virus nucleic acid detection reagents (Table 4), 2019-nCoV nucleic acid detection reagents national reference products (Table 5) and SARS plasmids were selected The carrier (from the State Key Laboratory of Respiratory Diseases, Guangzhou Medical University) was used to detect the kit.

[0131] Table 4 National Standards of Influenza Virus

[0132]

[0133] Table 5 National Reference Products of 2019-nCoV Nucleic Acid Detection Reagents

[0134]

[0135] Continuation

[0136]

[0137] experimental design:

[0138] Using some national reference materials for the second-generation influenza A / B nucleic acid detection reagents and 2019-nCoV nucleic acid detection reagents for the six respiratory virus nucleic acid detection kits (constant temper...

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Abstract

The invention relates to the field of medical detection, in particular to a primer probe combination for detecting six respiratory viruses, a kit and application. According to the kit provided by theinvention, a method of combining isothermal amplification and a micro-fluidic chip is adopted; the kit can accurately detect novel coronavirus (2019-nCoV) S and N target genes and influenza A virus, novel influenza A H1N1 virus (2009), influenza A H3N2 virus, influenza B virus and respiratory syncytial virus at the same time, and the purpose of distinguishing the novel coronavirus from common influenza virus is achieved. The kit disclosed by the invention has the advantages of multiple detection indexes, simplicity and convenience in operation, short detection time and the like, six virus indexes can be simultaneously detected by one-time sample adding, the time from sample treatment to report is within 1.5 hours, and the kit is more time-saving and labor-saving than qPCR method.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a combination of primers and probes, a kit and an application for detecting six respiratory viruses. Background technique [0002] The initial symptoms of 2019-nCoV patients are mostly fever, fatigue and dry cough, and gradually develop severe symptoms such as dyspnea. Most patients have a good prognosis, and some severe cases may develop acute respiratory distress syndrome or septic shock, and even die. Currently, there is no specific treatment for the disease. The incubation period of the disease is generally 3 to 7 days, the shortest incubation period is one day, and the longest incubation period is 14 days. The incubation period is contagious. Mainly through droplet transmission and contact transmission, there may be a risk of aerosol transmission in closed and unventilated places, and prevention and isolation treatment need to be strengthened. The population is generally...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2600/16Y02A50/30
Inventor 刘莹莹葛少林马银平牟海青韩春燕徐婷婷冯丽邢婉丽李为民程京
Owner CAPITALBIO CORP
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