Nucleic acid detection kit for synchronously identifying and diagnosing newcastle disease virus and avian influenza virus

Abstract

The invention belongs to the field of inspection and quarantine technology. Specifically, the invention is a nucleic acid detection reagent kit for synchronous discriminating and diagnosing avian influenza virus and newcastle disease virus. The detection reagents of the reagent kit include extraction reagent for extracting virus by silicon gel absorption column method, detection amplification reagent for detecting nucleic acid by RT-PCR Taq Man fluorescent probe method, and pretreatment liquid for solid tissue specimen for extracting virus RNA. Further, the invention employs in vitro transcription RNA as a positive contrast of the reagent kit. The reagent kit can rapidly and synchronously discriminate and diagnose avian influenza virus and newcastle disease virus which are highly infectious among avian plagues and have similar symptom, determine current major prevalent subtypes, such as H5, H7, H9, etc., and discriminate whether a infection source is an avian influenza having high pathogenicity, non-pathogenic avian influenza or mildly pathogenic avian influenza to human. The reagent kit is suitable for livestock and veterinarian station, import and export inspection and quarantine bureau, as well as other laboratories, and can be used for large-scale detection of influenza and epidemic surveillance.

Description

technical field [0001] The invention belongs to the technical field of inspection and quarantine, in particular to a nucleic acid detection kit for synchronous differential diagnosis of Newcastle disease virus and avian influenza virus. Background technique [0002] Avian influenza (European chicken plague, true chicken plague) and Newcastle disease (Asian chicken plague, false chicken plague) are both Class A severe poultry infectious diseases designated by the International Veterinary Bureau. They both have the characteristics of rapid spread, acute onset, and high mortality. It is also listed as a first-class animal disease in law. The symptoms of avian influenza (denoted as AIV) and Newcastle disease (denoted as DNV) are similar, the pathological process is indistinguishable, and the social hazards are very different: the current regulations on the control and culling of these two diseases are different. Highly pathogenic avian influenza such as H5N1 and H7 can break th...

AI Technical Summary

Problems solved by technology

[0004] There are many patent applications for avian influenza nucleic acid detection, but these applications are single-item detection, or multi-item PCR-electrophoresis detection, or multiple fluorescent PCR detection of H5, H7, and H9 subtypes, which is laborious and laborious, and cannot achieve Newcastle disease. Simultaneous identification and detection of avian influenza general type, H5, H7, H9, N1, etc.

There is only one Chinese patent application involving simultaneous detection of avian influenza and Newcastle disease, the application number is 200510042527.1 (name: method for detection of avian influenza and Newcastle disease virus by composite quantitative polymerase chain reaction), but it uses a non-probe technology but a fluorescent The disadvantage of the dye melting curve analysis method is that it has poor specificity and cannot type avian influenza; the reason is that the fluorescent dye melting curve analysis method with poor specificity cannot be used for multi-item research, otherwise too many amplification bands interfere and the Tm cannot be analyzed

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