MLPA (multiplex ligation-dependent probe amplification) identification kit capable of detecting multiple pathogens causing sudden death of pigs

Abstract

The invention relates to a biological detection technology and aims to provide an MLPA (multiplex ligation-dependent probe amplification) identification kit capable of detecting multiple pathogens causing sudden death of pigs. The kit comprises an RT (reverse transcription) and pre-amplification primer mixed liquid, a probe mixed liquid, an MLPA buffer solution, a ligation buffer solution A, a ligation buffer solution B, ligase Ligase-65, a PCR (polymerase chain reaction) mixed liquid, SALSA polymerase, negative control and positive control. Compared with the prior art, the kit has the characteristics of high throughput, good specificity and high sensitivity. The kit can detect 6 pathogens causing sudden death of pigs simultaneously and can detect 96 samples simultaneously each time by useof a full-automatic nucleic acid analyzer. Through probe design, sequence comparison and blast analysis, a probe is ensured to be only bound with a target gene. During detection, all that is requiredis to amplify a target fragment in a corresponding size from a corresponding template, and other pathogens are not amplified. The target gene is enriched through the added step of RT-PCR, and one copied target gene can be detected to the minimum.

Description

technical field [0001] The invention provides a multiplex connection probe amplification for detecting porcine reproductive and respiratory syndrome virus, swine fever virus, Nipah virus, African swine fever virus, Clostridium perfringens and Actinobacillus pleuropneumoniae The identification detection kit, primers and probes can simultaneously detect 6 kinds of pathogens that cause sudden death in pigs, and belong to the field of inspection and quarantine. Background technique [0002] Porcine reproductive and respiratory syndrome virus (PRRSV), swine fever virus (CSFV), Nipah virus (NiPV), African swine fever virus (ASFV), Clostridium perfringens and Actinobacillus pleuropneumoniae (Actinobacillus pleuropneumoniae) is a common pathogen causing sudden death syndrome in pigs. Although African swine fever and porcine Nipah disease have not been found in my country, the reports of existing diseases in neighboring countries and Asian regions should also be paid attention to, a...

AI Technical Summary

Problems solved by technology

Although there are corresponding methods and strategies for the factors that cause sudden death in pigs, there is no method that can identify multiple diseases that cause sudden death in pigs at one time

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