Triple rpa detection kit for bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus

Abstract

The invention discloses a triple RPA detection kit for bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus, comprising SEQ.ID.NO.1 and SEQ.ID.NO.5, SEQ.ID.NO. RPA of bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus composed of nucleotide sequences shown in .8 and SEQ.ID.NO.13 and SEQ.ID.NO.15 and SEQ.ID.NO.18 Detection primer set. The kit is based on the recombinase polymerase amplification method, uses cDNA formed by reverse transcription of RNA as a template to carry out amplification reaction, and uses nucleic acid gel electrophoresis to obtain detection results. The invention can be used to detect bovine viral diarrhea virus, bovine coronavirus and bovine rotavirus simultaneously, has simple operation, high sensitivity and specificity, and provides a rapid detection reagent for on-site screening of 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 the occurrence of cattle diarrhea, among which viral infection is the main reason. With the expansion of the scale of cattle raising, the incidence of diarrhea in calves is on the rise, and the harm 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 (BRV). [0003] BVDV belongs to Flaviviridae, Pestivirus, and its genome is single-stranded positive-stranded RNA. The natural host of BVDV includes pigs, cattle, sheep and other ruminants, which can cause fever, mucosal erosion, diarrhea, and r...

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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