PCR detection method for detecting four kinds of poultry respiratory disease pathogens simultaneously

Abstract

The invention belongs to the field of poultry disease detection and particularly relates to a one-step multiplex-PCR detection method for detecting four kinds of poultry respiratory disease pathogens simultaneously. The method is characterized by comprising the steps of providing a disease material possibly containing the four kinds of pathogens, extracting RNA, adding primers of the four kinds of pathogens to conduct PCR amplification and judging electrophoresis to obtain the result. The detection method is simple, convenient to operate and stable in accuracy and repeatability, whether the disease material contains the four kinds of pathogens can be confirmed in one experiment, 3 / 4 of detection cost is saved, detection time is shortened, and detection efficiency is improved.

Description

technical field [0001] The invention belongs to the field of poultry disease detection, in particular to a PCR detection method for simultaneously detecting four kinds of poultry respiratory disease pathogens. Background technique [0002] Avian influenza (Avian influenza, AI, including H5 subtype and H9 subtype), chicken Newcastle disease (Newcastle disease, ND), chicken infectious bronchitis (infectious bronchitis, IB) infection are four kinds that bring significant economic losses to the poultry industry. infectious disease. They have common respiratory symptoms clinically and can infect chickens, ducks, geese, pigeons and birds of different ages, causing high morbidity and mortality. Timely and accurate detection of these four pathogens is crucial for clinical disease prevention and control. [0003] Since the four pathogens are all RNA viruses, the traditional method requires a two-step reaction (reverse transcription and PCR) to obtain laboratory test results. The...

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Problems solved by technology

[0004] Due to the different reaction conditions, reverse transcription and PCR must be carried out in different reaction systems successively, increasing the number of experimental procedures and increasing the probability of experimental contamination

And because the optimal Tm values ​​of the primers required to amplify different pathogenic target fragments are different, traditional methods can only add one specific primer to the PCR reaction system each time for amplification, and only one pathogenic target fragment can be obtained at a time. Test results

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