PCR (Polymerase Chain Reaction) primer group and kit for jointly detecting various respiratory viruses

A detection primer and joint detection technology, applied in the biological field, can solve the problems of multiple detection links, complex and cumbersome operations, and low throughput, and achieve the effects of shortened detection time, accurate detection, and simple operation

Pending Publication Date: 2022-04-01
重庆巴斯德生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has the following disadvantages: (1) The operation is complicated and cumbersome, and it is not easy to standardize; (2) The cultivation period is long; (3) There are many detection links; (4) The technical requirements for personnel are relatively high

Method used

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  • PCR (Polymerase Chain Reaction) primer group and kit for jointly detecting various respiratory viruses
  • PCR (Polymerase Chain Reaction) primer group and kit for jointly detecting various respiratory viruses
  • PCR (Polymerase Chain Reaction) primer group and kit for jointly detecting various respiratory viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] When the examples give numerical ranges, it should be understood that, unless otherwise stated in the present invention, the two endpoints of each numerical range and any value between the two endpoints can be selected. 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. In addition to the specific methods, equipment, and materials used in the embodiments, according to those skilled in the art's grasp of the prior art and the description of the present invention, the methods, equipment, and materials described in the embodiments of the present invention can also be used Any methods, devices and materials of the prior art similar or equivalent to the practice of the present invention. The preparation and use method of embodiment 1 kit

[0064] The primers for detecting influenza A virus, the primers for influenza A virus H1N1 (2009), the primers for influenza A virus H...

Embodiment 2

[0077] The detection effect evaluation of embodiment 2 kit

[0078] The embodiment of the present invention provides a specific usage method of the kit. Each pair of detection primers shown in the sequence in Table 1, the pair of exogenous RNA internal reference amplification primers, and the OneStep RT-PCR Enzyme MIX purchased from QIAGEN Mix evenly to obtain multiple RT-PCR reactions, and the concentration of each primer in the multiple PCR reactions is shown in Table 3.

[0079] table 3

[0080]

[0081]

[0082] 1. Collect nasopharyngeal swab specimens from patients infected with human respiratory syncytial virus or influenza A virus H1N1 (2009) according to the standard operation. After collection, they should be placed in ice packs and sent for inspection immediately.

[0083] 2. After the sample is pre-treated, 10 μL of exogenous RNA internal reference is added to jointly extract the RNA in the sample to obtain the RNA sample to be tested.

[0084] 3. Preparation ...

Embodiment 3

[0099] Sensitivity and specificity analysis of embodiment 3 kit

[0100] sensitivity analysis:

[0101] Perform 10-fold serial dilutions on the positive quality control products of the seven virus detection targets, and the concentrations are 1×10 6 copies / mL, 1×10 5 copies / mL, 1×10 4 copies / mL, 1×10 3 copies / mL, 1×10 2 copies / mL and 10copies / mL, repeat 3-5 samples for each gradient dilution, use the same as in Example 2, determine the seven virus multiple PCR detection system, carry out multiple PCR amplification and fragment analysis detection, until If no signal was detected, 20 repeated tests were performed on each copy, and the positive detection rate level of 95% was used as the lowest lower limit of detection (LOD), which was the sensitivity.

[0102] The detection sensitivity of kit of the present invention, as shown in the following table:

[0103] Table 6

[0104]

[0105]

[0106] Specific analysis:

[0107] The specificity of the detection method esta...

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Abstract

The invention relates to the technical field of biology, in particular to a PCR (Polymerase Chain Reaction) primer group and a kit for jointly detecting various respiratory viruses. The PCR primer group comprises an influenza A virus detection primer pair, an influenza A virus H1N1 detection primer pair, an influenza A virus H3N2 detection primer pair, an influenza B virus detection primer pair, a human respiratory syncytial virus detection primer pair, a human metapneumovirus detection primer pair and a rhinovirus/enterovirus detection primer pair. The kit comprises a multiplex RT-PCR (Reverse Transcription-Polymerase Chain Reaction) reactant, and the multiplex RT-PCR reactant comprises the PCR primer group. The primer group and the kit disclosed by the invention can be used for jointly detecting and identifying seven respiratory viruses, the method is simple to operate, accurate in detection, high in sensitivity and high in specificity, the detection time is remarkably shortened, and a detection result can be obtained within 2-3 hours.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PCR primer set and a kit for jointly detecting multiple respiratory viruses. Background technique [0002] The morbidity, mortality, hospitalization rate, and medical burden of respiratory tract infection are high, and respiratory viruses such as influenza virus and human respiratory syncytial virus are the main causes, especially in young children, pregnant women, the elderly, and patients with other comorbidities. Influenza virus and human respiratory syncytial virus are also the main pathogens of acquired pneumonia. In addition to influenza virus and human respiratory syncytial virus, common clinical viruses that can cause respiratory tract infections include human metapneumovirus, rhinovirus, parainfluenza virus, coronavirus, adenovirus, boca virus, etc. [0003] At present, the commonly used respiratory virus detection methods mainly include the following: virus isolation and...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
Inventor 张祥林张劲松侯艳雯魏鹏胡秋萍
Owner 重庆巴斯德生物医药科技有限公司
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