SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis

Abstract

The invention relates to a SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis. The method provided by the invention comprises the following steps of: extracting pig Lawsonia Intracellularis DNA, designing primers, carrying out PCR amplification on aspa gene segments of PPE (polyphenylene oxide polyphenyl ether), cloning and identifying aspa genes of pig proliferative enteropathy, preparing a standard plasma template, amplifying fluorogenic quantitative PCR so as to obtain an amplification kinetic curve and a standard curve, testing the specificity, the sensitivity, the repeatability and stability, and detecting clinical disease material samples. The method provided by the invention has favorable specificity, sensitivity, repeatability and stability, and can identify and diagnose clinical suspected PPE infected or morbidity pigs simply, conveniently, rapidly, specifically, sensitively and accurately, and can detect and monitor the infection in the early stage.

Description

technical field [0001] The invention belongs to the technical field of bacterial detection, and in particular relates to a SYBR Green I fluorescent quantitative PCR detection method for Lawsonia intracellulare of porcine hyperplastic enteritidis. Background technique [0002] Porcine proliferative enteritis (PPE) is a contagious disease of pigs caused by Lawsonia intracellularis (LI). Lawsonia intracellulare is a curved obligate intracellular parasite, which mainly parasitizes the ileum, colon, and cecum of pigs and other animals, causing adenomatous hyperplasia of immature intestinal epithelial cells. At present, the disease has become a global epidemic. There are many recessively infected pigs, and the losses are easily overlooked. In recent years, China, Japan, South Korea and Taiwan in Asia have repeatedly occurred porcine proliferative enteritis. Sexual diarrhea, causing growth retardation of pigs, seriously reducing feed remuneration. In addition, the disease often h...

AI Technical Summary

Problems solved by technology

At present, the disease has become a global epidemic. There are many recessively infected pigs, and the losses are easily overlooked. In recent years, China, Japan, South Korea and Taiwan in Asia have repeatedly occurred porcine proliferative enteritis. Sexual diarrhea, causing growth retardation of pigs, seriously reducing feed remuneration

In addition, the disease often has mixed infection with other viruses and bacterial diseases, which makes the clinical symptoms more complicated, and it is difficult to make an accurate judgment, especially it is not easy to be associated with piglet paratyphoid, porcine transmissible gastroenteritis (TGEV) Diarrhea (PEDV) Diarrheal Disease Distinguished, etc.

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