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H5N1 type poultry grippal virus fluorescent augmentation detection kit and detection method

The technology of a bird flu virus and detection kit is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms. It can solve the problems of low separation rate, high requirements and limitations, and achieve simple and convenient sampling, high sensitivity and judgment. Convenient and accurate effect

Inactive Publication Date: 2010-12-15
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the virus isolation time is long, the requirements are high, and the isolation rate is low, and the double serum of the patient in the acute phase and the recovery phase is collected for antibody determination to confirm, and it cannot be diagnosed in time. Therefore, these two methods are greatly affected in use. limits

Method used

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  • H5N1 type poultry grippal virus fluorescent augmentation detection kit and detection method
  • H5N1 type poultry grippal virus fluorescent augmentation detection kit and detection method
  • H5N1 type poultry grippal virus fluorescent augmentation detection kit and detection method

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Embodiment 1

[0067] Utilize the TaqMan-MGB fluorescent quantitative RT-PCR method described in the present invention to detect 1 part of the lower respiratory tract aspirate specimen of the suspected H5N1 highly pathogenic avian influenza virus infection patient that occurred in Zhejiang Province in February 2006, and the result was positive (the The sample was re-examined by the Chinese Center for Disease Control and Prevention and confirmed positive for H5N1).

[0068] (1) Determination of standard curve:

[0069] The reaction system is as follows:

[0070] 5×RT-PCR buffer 5μL

[0071] Dimercaptothreitol 5mM

[0072] RNase inhibitor RNasin 0.1 unit / μL

[0073] Specific amplification upstream primer 0.48μM

[0074] Specific amplification downstream primer 0.48μM

[0075] Specific probe 0.64μM

[0076] The deoxynucleoside triphosphate mixture is:

[0077] dATP, dTTP, dCTP, dGTP each 200μM

[0078] Ampli TaqR DNA Polymerase 2.5 enzyme activity units / reaction

[0079] AMV reverse tr...

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Abstract

The present invention provides a kind of H5N1-type avian influenza virus fluorescence amplification detection kit and detection method for detecting avian influenza virus byusing said kit. Said kit includes the following several portions: H5N1-type avian influenza virus hemagglutinin gene standard product, fluorescence amplification detection reagent and DNA polymerase and reverse transcriptase. The fluorescence amplification detection reagent mainly includes one-step method RT-PCR buffer fluid, specific amplification primer and specific probe and deoxynucleoside triphosphate mixture. Said method can directly detect pathogen nucleic acid, and can detect specific gene containing H5N1-type avian influenza virus.

Description

(1) Technical field [0001] The invention relates to a H5N1 type avian influenza virus fluorescence amplification detection kit and a detection method. (2) Background technology [0002] Highly pathogenic avian influenza (HPAI) is an acute and severe respiratory infectious disease caused by H5N1 avian influenza virus. Its morbidity and mortality are extremely high. It is one of the main severe infectious diseases that endanger poultry. class of infectious diseases. In recent years, studies have found that the highly pathogenic avian influenza virus H5N1 can break through the racial barrier, infect humans and cause disease or even death. Since 2003, the number of deaths due to the infection of avian influenza H5N1 virus in Asia and my country has reached 64 people, making it one of the vicious infectious diseases that seriously endanger human health. Rapid mutation has posed a great threat to human health. Therefore, the establishment of rapid and accurate H5 avian influenza...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 卢亦愚严菊英冯燕茅海燕徐昌平葛琼
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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