Bovine parainfluenza type-3 virus detecting fluorescence quantitative PCR kit and application thereof

Abstract

The invention discloses a bovine parainfluenza type-3 virus detecting fluorescence quantitative PCR kit and an application thereof. The kit comprises: a) an RNA extraction solution, b) a reverse transcriptase reaction solution, c) reverse transcriptase, d) an RNA enzyme inhibitor, e) a primer and a TaqMan probe, f) a standard positive DNA template, and g) a fluorescence quantitative PCR reaction solution. The kit is characterized in that primer sequence is respectively a sense primer and an antisense primer: the size of an amplicon is 129 bp; in the sequence of a fluorescence probe, the 5' end of the probe is marked with a fluorescence emission group FAM while the 3' end is marked with a fluorescence quenching group TAMRA; the standard positive DNA template transforms a colon bacillus DH5 alpha by a pGEM-T carrier inserted with bovine parainfluenza type-3 virus conservative F protein coding zone 188 bp segment, plasmid is extracted after proliferation, an A260 ration is measured in an ultraviolet spectrophotometer, and the plasmid is diluted by 10 times of gradient. The result of the kit is more accurate, reliable, stable and repeatable. The kit can also be applied to the epidemiology investigation of bovine epidemic influenza and provide technical support for relevant fundamental researches, thus having quite broad application prospect.

Description

Technical field:

[0001] The invention relates to the field of biotechnology, more specifically to a fluorescent quantitative PCR kit for detecting bovine parainfluenza type 3 virus, and also relates to the use of the fluorescent quantitative PCR kit, which can efficiently and conveniently detect Quantitative detection and quality control of bovine blood products and bovine serum can also be used for epidemiological investigation of bovine parainfluenza, and can also provide technical support for related basic research, with a wide application prospect. Background technique

[0002] Bovine parainfluenza virus type 3 (PI-3) belongs to the genus Respirovirus of the family Paramyxoviridae, and is a segmented negative-strand RNA virus with a diameter of virions of 120 ~ 180nm, nucleocapsid helical symmetry, containing hemagglutinin and neuraminidase activity. Serological studies have shown that the presence of PI-3 virus antibodies in healthy bovine serum is more common, so it i...

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