Triple RPA (recombinase polymerase amplification) detection kit for bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus

Abstract

The invention discloses a triple RPA (recombinase polymerase amplification) detection kit for a bovine viral diarrhea virus, a bovine coronavirus and a bovine rotavirus. The triple RPA detection kit comprises RPA detection primer groups for the bovine viral diarrhea virus, the bovine coronavirus and the bovine rotavirus, wherein the RPA detection primer groups are respectively composed of nucleotide sequences shown as SEQ ID NO.1 and SEQ ID NO.5, SEQ ID NO.8 and SEQ ID NO.13, and SEQ ID NO.15 and SEQ ID NO.18. The kit is based on a recombinase polymerase amplification method, cDNA formed by RNA reverse transcription is used as a template for amplification reaction, and a detection result is obtained by nucleic acid gel electrophoresis. The kit can be used for simultaneously detecting the bovine viral diarrhea virus, the bovine coronavirus and the bovine rotavirus, is simple to operate, has relatively high sensitivity and specificity, and provides a rapid detection reagent for on-site screening of the bovine diarrhea virus.

Description

technical field [0001] The invention relates to the field of viral nucleic acid detection, in particular to a triple RPA detection primer set, kit and detection method for bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus. Background technique [0002] Diarrhea is a common disease in cattle. There are many reasons for cattle diarrhea, among which viral infection is the main reason. With the expansion of cattle raising scale, the incidence of calf diarrhea is on the rise, and the damage is the most serious. As far as the current situation is concerned, the viruses that cause bovine diarrhea mainly include bovine viral diarrhea virus (BVDV), bovine coronavirus (BCoV) and bovine rotavirus (Bovine rotavirus, BRV). [0003] BVDV belongs to Flaviviridae and Pestivirus. Its genome is single-stranded positive-sense RNA. The natural hosts of BVDV include pigs, cattle, sheep and other ruminants. It can cause fever, mucosal erosion, diarrhea, and reproductive disor...

AI Technical Summary

Problems solved by technology

[0006] At present, there are only reports on the application of RPA technology to detect a single type of bovine diarrhea virus, for example, Chinese patents CN104894118A and CN107974513A involving the detection of BVDV, and Chinese patent CN108546779A involving the detection of BRV, while the detection of BCoV has not yet been reported involving RPA technology

Compared with PCR, RPA has more stringent requirements on primer design, which cannot be realized completely according to software design. Screening effective specific amplification primers and reaction conditions is a difficult problem in realizing multiple RPA detection.

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