Reverse transcription loop-mediated isothermal amplification (RT-LAMP) kit for porcine kobuvirus and application thereof

Abstract

The invention discloses a visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) kit for the porcine kobuvirus and application thereof. The kit comprises an RT-LAMP primer, 2*reaction buffer, effective microorganisms (EM), a fluorescent visual detection reagent, ultrapure water and a porcine kobuvirus ribonucleic acid (RNA) template, wherein the RT-LAMP primer comprises outer primers F3 and B3, inner primers FIP and BIP and loop primers LF and LB. The kit is applied to detecting the pathological tissues of the porcine kobuvirus and virus-containing excrement. Specific detection and sensitivity detection verify that the RT-LAMP kit provided by the invention can monitor reaction in real time and quantitatively detect the copy number of the porcine kobuvirus, quickly and accurately obtain the detection results and bring convenience for simply, conveniently, quickly and reliably detecting the porcine kobuvirus.

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 porcine ridge virus reverse transcription loop-mediated isothermal amplification kit and its application. Background technique [0002] Porcine crest virus (Kobuvirus, Kobu) is a single-stranded positive-sense RNA virus belonging to the family Picornaviridae and belonging to the genus Crestavirus. The diameter of the virus particle is about 30nm, and it is icosahedral and symmetrical, without a capsule. In 1998, a new type of RNA virus was found in the stool samples of patients with gastroenteritis in Aichi Prefecture, Japan. It was named Aichivirus according to the place of discovery. In 1997, through genetic analysis, it was confirmed that Aichivirus was a new type of RNA virus. microRNA virus. Because the virion is rugged and ridge-shaped under the electron microscope, it is named cristaevirus. At presen...

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Problems solved by technology

[0004] The purpose of the present invention is to solve the problems such as difficulty in detection of porcine crest virus at the grassroots level, time-consuming and expensive equipment, and provide a simple, fast and accurate detection kit for the basic level of porcine crest virus, the technical solution used to achieve the purpose of the present invention It is: a porcine ridge virus reverse transcription loop-mediated isothermal amplification kit, the kit includes RT-LAMP primers, 2× reaction buffer, EM, fluorescent visual detection reagents, ultrapure water and porcine ridge virus RNA template; The RT-LAMP primers include outer primers F3 (SEQ ID NO: 1) and B3 (SEQ ID NO: 2), inner primers FIP (SEQ ID NO: 3) and BIP (SEQ ID NO: 4) and loop primers LF (SEQ ID NO: 5) and LB (SEQ ID NO: 6);

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