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Kit for detecting H1N1 influenza A virus by real-time fluorescence RT-PCR

A RT-PCR, real-time fluorescence technology, used in fluorescence/phosphorescence, microorganism-based methods, and microbial determination/inspection to achieve good sensitivity, improve reliability and accuracy, and avoid false negatives and false positives.

Active Publication Date: 2010-11-17
DAAN GENE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, no real-time fluorescent RT-PCR kit for detecting influenza A H1N1 virus has been developed at home and abroad, so the development of this kit has become a top priority

Method used

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  • Kit for detecting H1N1 influenza A virus by real-time fluorescence RT-PCR
  • Kit for detecting H1N1 influenza A virus by real-time fluorescence RT-PCR
  • Kit for detecting H1N1 influenza A virus by real-time fluorescence RT-PCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: the development of type A H1N1 influenza virus detection reagent

[0042] 1. Design of primers and probes: through sequence comparison and analysis of the nucleic acid sequences of the reported influenza A (H1N1) viruses, the H1 subtype HA gene and the N1 subtype NA gene of the influenza A (H1N1) virus were respectively used as amplification targets According to the basic principles of primer and probe design, multiple pairs of primers and probes are designed by software and manually.

[0043] 2. Selection of clinical samples: According to relevant domestic and foreign literature reports, throat swabs, nasopharyngeal secretions and other respiratory samples as well as serum samples can be used

[0044] 3. Establishment and optimization of the reaction system

[0045] Sample preparation: use the constructed plasmid containing the target amplification region as the positive quality control product for the detection of influenza A (H1N1) virus; Respiratory sy...

Embodiment 2

[0054] Embodiment 2: Influenza A (H1N1) virus detection kit and its use

[0055] 1. Prepare a kit including the following components: 1 tube of RNA extraction solution (50ml / tube), 1 tube of H1 subtype primer-probe mixture (50 μl / tube), 1 tube of N1 subtype primer-probe mixture (50 μl / tube) ) 1 tube, RT-PCR reaction solution (720μl / tube) 1 tube, RT-PCR enzyme system (150μl / tube) 1 tube, negative quality control (200μl / tube) 1 tube, H1 subtype positive quality control ( 100μl / tube) 1 tube, N1 subtype positive quality control (100μl / tube) 1 tube.

[0056] 2. Specimen collection, transportation and storage

[0057] Specimens will be collected by clinicians according to the actual situation. Testable specimens include throat swabs, nasopharyngeal secretions, and serum. The collection methods are as follows: ① nasopharyngeal secretions: natural expectoration or induced sputum suction (such as positive oxygen pressure method, disposable baby suction tube method, and nebulized stea...

Embodiment 3

[0072] Example 3: Application of Influenza A (H1N1) Influenza Virus Detection Kit to Detect Samples

[0073] With positive quality control substance, negative quality control substance, specific reference substance in embodiment 1 as samples to be tested, detect these samples respectively according to the method of embodiment 2, positive quality control substance detection result is positive, negative quality control substance and The test results of specific reference products were all negative (see attached figure 1 ,2).

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Abstract

The invention relates to a kit for detecting an H1N1 influenza A virus by real-time fluorescence RT-PCR, in particular to a kit for early and rapidly diagnosing infection of the H1N1 influenza A virus by one-step real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) technology. The kit has high sensitivity and specificity. The kit of the invention realizes early and rapid diagnosis of the H1N1 influenza A virus in respiratory tract samples such as nasopharyngeal swabs, nasopharyngeal secretions and the like and samples such as blood serum and the like.

Description

technical field [0001] The present invention relates to a kind of kit for real-time fluorescence RT-PCR detection type A (H1N1) influenza virus, in particular to early and rapid diagnosis of type A (H1N1) influenza virus by one-step real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR) technology Infection kit. The kit has high sensitivity and specificity, and the kit can realize early rapid diagnosis of influenza A (H1N1) virus in throat swabs, nasopharyngeal secretions and other respiratory samples, as well as serum samples. Background technique [0002] Influenza is called flu for short, is an acute respiratory infectious disease caused by three kinds of influenza viruses A, B and C. Influenza A viruses can be divided into many subtypes according to their surface (H and N) structures and their genetic characteristics. So far, influenza A viruses have found 16 subtypes of hemagglutinin (H1-H16), neuramine There are 9 subtypes of acidase (N1-N9)....

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64C12R1/93
Inventor 李明高秀杰陈华云何蕴韶
Owner DAAN GENE CO LTD
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