RT-PCR (Reverse Transcription-Polymerase Chain Reaction) discrimination and detection method of virulent classical swine fever virus (CSFV) Shimen strain and attenuated vaccine C strain

Abstract

The invention discloses a RT-PCR (Reverse Transcription-Polymerase Chain Reaction) discrimination and detection method of a virulent classical swine fever virus (CSFV) Shimen strain and an attenuated vaccine C strain. The method comprises the following steps of: (1) synthesizing primers, wherein a CSFV virulent detection primer is CSFV-S-F / SCFV-S-R, and a CSFV attenuated detection primer is CSFV-C-F / CSFV-C-R; (2) extracting a virus RNA (Ribose Nucleic Acid) and synthesizing a cDNA (Complementary Deoxyribose Nucleic Acid) template; and (3) carrying out PCR amplification so that one 256 bp strip can be amplified from the attenuated template and one 428 bp strip can be amplified from the virulent strip. By using the method, only once PCR is needed after the reverse transcription; the method is suitable for a general laboratory to develop discrimination and detection of the virulent classical swine fever virus and the attenuated vaccine; and the immune uninfected pigs can be prevented from being killed by mistake so that certain economic benefits can be generated.

Description

technical field [0001] The invention relates to a RT-PCR differential detection method for CSFV virulent (CSFV) Shimen strain and vaccine attenuated C strain, belonging to the field of molecular biology technology. Background technique [0002] Classical swine fever (CSF) or (Hog cholera, HC) is a severe viral infectious disease that seriously harms pigs. Because CSF is very harmful to the pig industry and is difficult to control effectively, the World Organization for Animal Health (OIE) once listed it as a class A animal disease, and listed it as a must in 1.2.3.5 of the 2008 version of the "Terrestrial Animal Code" One of the 7 important infectious diseases of pigs reported. In 2008, my country's Ministry of Agriculture Announcement No. 1125 "List of Types of Animal Epidemic Diseases of the First, Second, and Third Types" also listed it as one of 17 kinds of first-class animal infectious diseases. All countries in the world have adopted strict prevention and control mea...

AI Technical Summary

Problems solved by technology

Although attenuated vaccine immunized pigs can produce complete immune protection, there is also the problem that CSFV naturally infected pigs and vaccine-immunized pigs cannot be distinguished serologically, which brings great difficulties to the identification and detection of CSFV-infected pigs and vaccine-immunized pigs; The new swine fever vaccine developed in recent years has its advantages in preventing the virulence of the attenuated vaccine from returning to strength, but there are also problems in clinical application. For example, various swine fever subunit vaccines have low immune efficiency and cannot provide complete immune protection. problems, and cannot meet the urgent immunization needs of CSFV infection

Especially in the late 1970s, the prevalence and incidence characteristics of domestic swine fever changed greatly, and the clinical manifestations tended to become more complicated. So-called "atypical swine fever", "mild swine fever" and "poisonous swine fever" appeared Porcine Syndrome", the number of typical cases decreased, making it difficult to diagnose the disease

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