Universal RT-PCR (Reverse Transcription Polymerase Chain Reaction) detection primer and detection method for avian pneumovirus

Abstract

The invention discloses a universal RT-PCR (Reverse Transcription Polymerase Chain Reaction) detection primer and a detection method for avian pneumovirus (APV), belonging to the technical field of detection for avian viruses.. The universal RT-PCR detection primer has nucleotide sequences shown as SEQ ID NO:1 to SEQ ID NO:2. The detection method is as follows: the universal RT-PCR detection primer for avian pneumovirus and the RNA (Ribonucleic Acid) of a sample to be detected are subjected to RT-PCR, and the presence of avian pneumovirus in the sample to be detected is confirmed if the PCR product fragment is 319bp o 320bp. The method disclosed by the invention is high in primer specificity, small in fragment and high in sensitivity, can finish the entire PCR process within 2 hours and can also specifically detect the presence of various subtype APVs, thus greatly reducing the erroneous diagnosis for APV infection.

Description

Technical field [0001] The present invention involves the field of poultry virus detection technology, and specifically involves a pair of poultry pulmonary virus common RT-PCR detection primers and detection methods. Background technique [0002] Avian Pneumovirus (APV), also known as Turkey Rhinot Racheitis Virus (TRTV), belongs to the genus of sub -chipocarium virus, which is a single -chain negative RNA virus, can cause fireChicken and some other poultry upper respiratory diseases.The APV genome contains eight major structural protein genes, with a total length of the genome of about 13.4 kb, and the order of eight genes from 3 'to 5' end is N-P-M-F-M2-SH-G-L.At first, people thought that APV had only one serum type, two gene subtypes (A and B), which can effectively distinguish between the two genotypes through nucleotide sequence analysis and monoclonal antibody analysis.In the late 1990s, two different new types of APV strains (type C and D) appeared in the United States a...

AI Technical Summary

Problems solved by technology

However, using subtype-specific RT-PCR as the preferred detection technology will lead to often being ignored when there is a new type of APV, which is not conducive to the effective detection of APV infection. D-type APV has existed in France for more than 15 years. recognized

Existing primers for APV detection can only detect one of the subtypes. In order to facilitate the comprehensive detection of the virus, a universal RT-PCR detection method is urgently needed

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