H7 subtype bird flu virus RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification) test kit and application thereof

A RT-LAMP, avian influenza virus technology, applied in the direction of determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of complex detection instruments, limitations, etc., to achieve less sample, good specificity, effective early and rapid The effect of the diagnostic approach

Inactive Publication Date: 2011-07-27
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, complex testing instruments (such as PCR instrument, fluorescent quantitative PCR i...

Method used

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  • H7 subtype bird flu virus RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification) test kit and application thereof
  • H7 subtype bird flu virus RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification) test kit and application thereof
  • H7 subtype bird flu virus RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification) test kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0041] 1 Test materials and methods

[0042] 1.1 Virus strains and reagents

[0043] The standard reference strain of avian influenza virus H1~H15 subtype used in this test (purchased from China Veterinary Drug Control Institute);

[0044] H7 subtype avian influenza virus (HPAIV (A / FPV / Rostock / 1934 (H7N1)) and LPAIV (A / Chicken / HeB / 2 / 02 (H7N2)) and its Newcastle disease virus (NDV), chicken infectious bronchitis virus (IBV), chicken infectious bursal virus (IBDV), chicken Marek's disease virus (MDV), chicken infectious laryngotracheitis virus (ILTV), chicken infectious anemia virus (CIAV), chicken leukemia virus (ALV) (purchased from China Veterinary Drug Control Institute).

[0045] RNA Amplification Kit (RT-LAMP) and Fluorescent Detection Reagent were purchased from Japan Eiken Company; RNA Both Total RNA Isolation system and Access RT-PCR system were purchased from Promega. 10-day-old SPF chicken embryos and 6-week-old SPF chickens were provided by the SPF Experimental ...

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Abstract

The invention discloses an H7 subtype bird flu virus RT-LAMP (Reverse Transcriptase Loop-Mediated Isothermal Amplification) test kit and application thereof. The H7 subtype bird flu virus RT-LAMP test kit comprises a Bst DNA, a Bst buffering liquid, AMV (Alfalfa Mosaic Virus) reverse transcriptase, dNTPs, betaine, magnesium sulfate, fluorescent dye, an RT-LAMP primer and distilled water, wherein the RT-LAMP primer consists of a pair of external primers, a pair of internal primers and a pair of ring primers, wherein the pair sequences of the external primers are shown in the SEQ ID No.1 and SEQ ID No.2; the pair sequences of the internal primers are shown in the SEQ ID No.3 and the SEQ ID No.4; and the pair sequences of the ring primers are shown in the SEQ ID No.5 and the SEQ ID No.6. The H7 subtype bird flu virus RT-LAMP test kit has the characteristics of small amount of samples, high specificity, high sensitivity, fastness and accuracy and the like, and provides a simple, convenient and effective early quick testing way for a prevention, control and early warning mechanism of H7 subtype bird flu.

Description

technical field [0001] The invention relates to an avian influenza virus detection kit, in particular to an H7 subtype avian influenza virus RT-LAMP detection kit and an application thereof, belonging to the field of avian influenza virus detection. Background technique [0002] The outbreak and prevalence of avian influenza have caused huge economic losses to the animal husbandry and posed a serious threat to human health. Among the HA subtypes of avian influenza, only the H5 and H7 subtypes are highly pathogenic to birds (Alexander DJ, 1995), which are called highly pathogenic avian influenza (HPAI). It caused 100% death of chicken flocks and was identified as Class A infectious disease by OIE (Truszczynsky et al, 2000). The natural hosts of influenza A virus are wild waterfowl, gulls and shore birds, and domestic ducks are also a huge reservoir of AIVs, and are also considered to be an important intermediate host for the transmission of AIVs from natural hosts to poultry...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 包红梅王秀荣陈化兰李雁冰田国彬熊永忠
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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