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H7N9 influenza virus A detection kit and detection method

A technology of influenza A virus and detection kits, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve problems such as long cycle time, low positive rate, and high laboratory hardware requirements

Active Publication Date: 2013-08-14
北京鑫诺美迪基因检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The "gold standard" for influenza virus detection is virus culture, but the virus culture operation is complicated, the laboratory hardware requirements are high, the cycle is long, and the positive rate is low, which limits its application.
Currently there is no ready-made rapid detection kit, this article describes a rapid detection kit for H7N9 virus

Method used

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  • H7N9 influenza virus A detection kit and detection method
  • H7N9 influenza virus A detection kit and detection method
  • H7N9 influenza virus A detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] The kit includes four indicators, that is, one sample contains four tubes for detection, and the concentration, volume and nucleotide sequence of the primers and probes are as follows:

[0070] 1. System (1): Universal detection of influenza A virus

[0071] The specific composition of influenza A virus universal detection primers and probes is shown in Table 1

[0072] Table 1 Universal Detection Primer and Probe Concentration and Volume

[0073] join in turn

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Amount for 1 test

①

Infa-F1(W)(10pmoL / μL)

1.5μL

②

Infa-R1(W)(10pmoL / μL)

1.5μL

③

Infa-FP1(W) (2.5pmoL / μL)

1.5μL

④

RNaseP-F1 (10pmoL / μL)

0.5μL

⑤

RNaseP-R1 (10pmoL / μL)

0.5μL

⑥

RNaseP-FP1 (2.5pmoL / μL)

0.5μL

[0074] sequence

[0075] Table 2 Nucleotide sequences of viral target gene primers and probes in the universal detection system

[0076] Infa-F1(W)

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experiment example

[0140] In order to investigate the specificity and sensitivity of the detection kit in Example 1 according to the method of Example 2, the following experiments were carried out.

Embodiment ( 1

[0142] Select 2 cases of 2013H7N9 avian influenza virus, 1 case of non-2013H7N9 type avian influenza virus, 1 case of influenza A H7N2 virus, 1 case of influenza A H1N1 (2009) virus, 1 case of influenza A H3N2 virus, and 1 case of seasonal H1N1 Throat swab samples for influenza virus testing. In order to verify the specificity of the detection method, samples of influenza B virus, respiratory syncytial virus, parainfluenza virus, adenovirus, coronavirus, and enterovirus EV71 / CA16 were collected to verify the specificity of the method.

[0143] The detection method is as described above, and the detection results are as follows:

[0144] (1) The four detection indicators of the 2013H7N9 avian influenza virus sample, namely MP, HA, NA, and NP, were all positive.

[0145] (2) Non-2013H7N9 avian influenza virus samples were positive for MP, HA, and NA, and negative for NP.

[0146] (3) MP and HA of influenza A (H7N2) virus samples were positive, and NA and NP were negative.

[...

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Abstract

The invention discloses an H7N9 influenza virus A detection kit and detection method. In the method, H7N9 influenza virus MP, HA, NA and NP genes are respectively selected for designing a specific primer and a probe, and after the sample RNA (ribonucleic acid) nucleic acid is purified, the fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) technology is used for performing qualitative detection on the virus RNA, so as to affirm the influenza virus and primary type of the influenza virus and finally determine whether the influenza virus belongs to HTN9 type. The kit and the detection method disclosed by the invention, are simple to operate, quick in result acquisition and accurate in detection result, provide practical means for rapid detection on H7N9 influenza virus A, and have positive social value.

Description

technical field [0001] The invention relates to a detection method and a detection kit for H7N9 virus. Background technique [0002] From March to April 2013, a bird flu outbreak occurred in many places in East my country, killing many people and gradually spreading to many places. As of May 1, 127 cases of H7N9 bird flu have been confirmed and 26 people have died, involving Beijing, Shanghai, Jiangsu, Zhejiang, Anhui, Jiangxi, Hunan, Shandong, Fujian and Henan provinces and cities. Studies have confirmed that the pathogenic source of the outbreak is a new type of recombinant H7N9 avian influenza virus. At present, they are all sporadic cases, and no epidemiological association has been found between the cases. The source of infection is still unclear. According to past experience and epidemiological investigation of this case, it is speculated that it may be birds carrying H7N9 avian influenza virus and their secretions or excretions. The virus is transmitted through the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 叶锋刘明坤迟磊宋玉亮
Owner 北京鑫诺美迪基因检测技术有限公司