Reverse transcription loop-mediated isothermal amplification kit for detecting pestedes petits ruminants viruses

Abstract

The invention discloses a reverse transcription loop-mediated isothermal amplification (RT-LAMP) kit for detecting pestedes petits ruminants viruses (PPRVs). The kit comprises an RT-LAMP primer, a 2X reaction buffer solution, an enzyme mixture (EM), a fluorescent visual detection reagent, ultrapure water and a reaction template, wherein the RT-LAMP primer includes outer primers F3 and B3, inner primers FIP and BIP and loop primers LF and LB. The detection kit is mainly used for detecting whether the PPRVs exist in lesion tissues suspiciously infected with the PPRVs. Specific detection and sensitivity detection prove that the RT-LAMP kit provided by the invention can monitor a reaction in real time and quantitatively detect a copy number of the PPRVs, a detection result is obtained quickly and accurately, and the convenience is brought for simply and quickly detecting the PPRVs.

Description

technical field [0001] The invention relates to the technical field of microbial detection, in particular to a rapid, visualized and real-time quantitative detection of Peste des petits ruminants virus (PPRV) reverse transcription loop-mediated isothermal amplification kit and its application. Background technique [0002] Peste des petits ruminants (PPR) is an acute, virulent, contagious disease caused by Peste des petits ruminants virus (PPRV), which mainly infects small ruminants such as goats and sheep, especially goats are highly susceptible, wild animals Occurs by chance, and the infection rate and mortality rate are as high as 100% and 90%. OIE once regarded the case as a Class A animal infectious disease that must be reported, and my country listed it as a Class I animal disease. Once discovered, urgent and severe measures must be taken to control and eradicate it. Large-scale outbreaks of epidemics in Laos, Bangladesh, India, Nepal, Russia, Pakistan, Myanmar and ot...

AI Technical Summary

Problems solved by technology

Since Peste des petits ruminants virus and rinderpest virus of the same genus have a common antigen in serum detection, the neutralization test also has a certain degree of cross-reaction, so the two are easily confused; competition ELISA is also time-consuming and requires the presence of live virus

Viral nucleic acid detection methods are mainly common RT-PCR method and fluorescent RT-PCR method. The detection sensitivity of the latter is higher than that of the former. Although these two methods are faster and more accurate than virus isolation and identification and serological detection methods, RT-PCR requires expensive PCR instrument, and agarose gel electrophoresis is required for the judgment of the results, which may easily cause laboratory contamination and lead to false positive results. More expensive than ordinary PCR machines, and the cost of reagents is higher, which is not conducive to grass-roots detection and on-site batch detection

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