Reagent kit for detecting peste des petits ruminant vaccine poison by pyrophosphoric acid sequencing technology

Abstract

The invention provides a reagent kit for detecting peste des petits ruminant vaccine poison by a pyrophosphoric acid sequencing technology. The reagent kit comprises a peste des petits ruminant specific primer and a pyrophosphoric acid sequencing primer, and the nucleotide sequences of the peste des petits ruminant specific primer and the pyrophosphoric acid sequencing primer are respectively shown as SEQ ID No. 2, SEQ ID No. 3 and SEQ ID No. 4. The RNA of a to-be-tested sample is extracted, and RT-PCR amplification is carried out by the specific primer. If the length of a primer-amplified fragment is 180bp, pyrophosphoric acid sequencing reaction is carried out. If a sequencing fragment is identical to an N gene target fragment (SEQ ID NO.1), peste des petits ruminant vaccine poison is determined. If at least one basic group of the sequencing fragment is different from the target fragment, the sample is judged as a negative one. The reagent kit can be used for the rapid detection of the peste des petits ruminant vaccine poison and has the characteristics of high throughout and low cost. A product can be directly used for sequencing, secondary treating is not required, the operation is very simple, the quantity of required sample is small, and the application prospects are great.

Description

technical field [0001] The invention relates to pyrosequencing technology, in particular to a kit for detecting the virus of Peste des petits ruminants vaccine by using the pyrosequencing technology. Background technique [0002] Peste des petits ruminants (PPR) is a highly contagious animal disease caused by Peste des petits ruminants (PPRV) of the genus Morbillivirus in the Paramyxoviridae family. The disease mainly infects domestic and wild small ruminants, and goats are especially susceptible. Its pathogenicity and molecular characteristics are very similar to rinderpest (RP), and it is defined as a category A animal disease by the World Organization for Animal Health (OIE). The foreign trade economy of countries affected by the disease, especially in Africa, the Middle East, the Near East and South Asia, has brought huge losses. Since the outbreak of the disease, it has been spreading to all parts of the world, posing a greater threat to countries without the disease, ...

AI Technical Summary

Problems solved by technology

In recent years, there have been many reports on the diagnosis of PPRV and the exclusion of rinderpest virus (RPV) interference by PCR, RT-PCR, ELISA and other methods, but there are few reports on the method of distinguishing and detecting PPRV vaccine strains from virulent strains

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