Multiplex-PCR (polymerase chain reaction) kit for detecting six viruses of sheep and goats simultaneously
A kit and multiple technology, applied in the field of multiple PCR detection kits, can solve the problems of difficult to distinguish in clinical diagnosis, slow growth and development of lambs, economic losses of sheep farming and other problems, and achieve short detection time, simple detection process and high sensitivity Effect
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Embodiment 1
[0027] Example 1: Multiplex PCR kit for simultaneous detection of six viruses carried by sheep and goats
[0028] The kit includes six pairs of specific primers, positive control, negative control, reverse transcription solution, PCR mixture and deionized water. Among them, the primers used for the detection of bluetongue virus were BTV-F and BTV-R; the primers used for the detection of foot-and-mouth disease virus were FMDV-F and FMDV-R; the primers used for the detection of Peste des Petits Ruminants virus were PPRV -F and PPRV-R; the primers used for sheep pox virus detection are SPPV-F and SPPV-R respectively; the primers used for goat pox virus detection are GTPV-F and GTPV-R respectively; the primers used for sheep pox virus detection The primers are ORFV-F and ORFV-R respectively;
[0029] The primers used for bluetongue virus detection are designed for the NS3 gene of bluetongue virus, and the primer sequences are BTV-F and BTV-R, respectively.
[0030] BTV-F: 5'-AGT...
Embodiment 2
[0048] Example 2: Rapid detection of bluetongue virus, foot-and-mouth disease virus, Peste des Petits Ruminants virus, sheep pox virus, goat pox virus and sheep aphthous disease virus
[0049] 1. Materials
[0050] (1) Positive\negative control
[0051] Positive controls: plasmids carrying the NS3 gene of bluetongue virus, plasmids carrying the 3D gene of foot-and-mouth disease virus, plasmids carrying the N gene of Peste des petits ruminants vaccine strain, plasmids carrying the whole gene of sheep pox virus 119555-119711, and plasmids carrying the whole gene of goat pox virus The plasmids of the 110977-111368 segment of the gene and the plasmid mixture carrying the 011 gene of sheep mouth disease virus were constructed and preserved by the Veterinary Microbiology Laboratory of the School of Veterinary Medicine, Northwest Agriculture and Forestry University. Ltd Synthetics.
[0052] Negative control: deionized water without PPRV, BTV, FMDV, SPPV, GTPV, and ORFV.
[0053] (...
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