CSFV detection method utilizing realtime fluorescence quantitative RT-PCR

Abstract

The invention discloses a CSFV (classical swine fever virus) detection method utilizing realtime fluorescence quantitative RT-PCR. According to results of singularity and sensitivity teats of a method established in the invention, the method can well distinguish the CSFV from FMDV (Foot And Mouth Disease Virus), TGEV (Transmissible Gastroenteritis Virus), PRRSV (Porcine Respiratory and Reproductive Syndrome Virus) and PCVII( Porcine Circovirus II), and sensitivity can reach an order of magnitude of 10 copy number. In addition, whole reaction process of the method established by the invention is operated in a close pipe, so as to reduce solution to the system at most. Compared with a traditional method, the method in the invention has a substantially shortened detection time, on the premise of guaranteeing detection result. For the whole process, comprising steps of obtaining of sample to be checked, nucleic acid extraction, preparation of fluorescence quantitative RT-PCR reaction system, and appearance of reaction result, can be finished within 2.5 h.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a method for accurate, specific and rapid detection of CSFV. Background technique [0002] Swine fever (swine fever, SF), also known as hog cholera (hog cholera, HC) and intestinal rotten fever, is called classical swine fever (classical swine fever, CSF) in Europe. Important infectious diseases once brought serious economic losses to the pig industry. [0003] The traditional diagnostic methods for CSF mainly include animal test, immunofluorescence test, serum neutralization test, ELISA, and monoclonal antibody test. Animal test is a sensitive method to detect CSFV, but the detection time is relatively long and most of the currently circulating CSFVs are low-virulence virus strains and do not show typical symptoms of CSF even after inoculation; the immunofluorescence test method is relatively specific, but In the actual detection application process, this technology requires...

AI Technical Summary

Problems solved by technology

Animal test is a sensitive method to detect CSFV, but the detection time is relatively long and most of the currently circulating CSFVs are low-virulence virus strains and do not show typical symptoms of CSF even after inoculation; the immunofluorescence test method is relatively specific, but In the actual detection application process, this technology requires fluorescent antibodies, fluorescent microscopes and inspectors with high detection technology; serum neutralization test and ELISA test are difficult to distinguish vaccine antibodies from antibodies produced after infection, except for a long detection time , it is prone to false positives; monoclonal antibody technology has strong specificity, but the operation process is complicated and the cost is high, and it is not suitable for the requirements of large-scale sample diagnosis

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