Taqman real-time fluorescence PCR (polymerase chain reaction) kit for detecting hog umbilical cord blood hog cholera viruses and application of kit

Abstract

The invention discloses a Taqman real-time fluorescence PCR (polymerase chain reaction) kit for detecting hog umbilical cord blood hog cholera viruses and an application of the kit. The kit comprises a pair of primers and a specific fluorescence probe, the sequence of the primer is shown as SEQ ID NO: I, and the sequence of the fluorescence probe is shown as SEQ ID NO: 3. The kit can specifically diagnose the hog umbilical cord blood hog cholera viruses, has high sensitivity and universality and excellent repeatability, can accurately evaluate and diagnose the sow cholera virus carrying elimination and piglet infection, can perform hog cholera purification, effect evaluation and the like and has a quite extensive application value. Besides, the kit needs simple instrument equipment in the detection process, is simple and convenient in detection process, short in detection time, not easy to pollute, low in technical operation requirement and easy to standardize and can be extensively popularized on a base layer, results are reliable, and quality can be easily controlled.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis, in particular to a Taqman real-time fluorescent PCR kit for detecting porcine umbilical cord blood swine fever virus and its application. Background technique [0002] Classical swine fever is a highly contagious and acute infectious disease of pigs caused by classical swine fever virus (CSFV). Sexual spotting, multiple hemorrhage, necrosis and infarction in internal organs, with very mild infectivity and high fatality rate. Early diagnosis of classical swine fever was based on typical clinical symptoms and pathological changes. In recent years, atypical clinical manifestations of classical swine fever are more common, such as reproductive disorders in sows, congenital infection of newborn piglets, persistent infection, immune tolerance and asymptomatic Toxic and other forms. It brings great challenges to the differential diagnosis of swine fever. [0003] With the development of b...

AI Technical Summary

Problems solved by technology

However, there are still few commercial kits independently developed in China

At the same time, the ordinary PCR detection method has the defects of prone to false positives, long operation time, nested amplification, and high technical requirements for technical operation.

[0004] The existing technology has the following deficiencies: ① Serological detection technology is difficult to evaluate the immune tolerance phenomenon in pigs, and it is relatively unable to reflect the antibody level of pigs more effectively, intuitively and accurately; ② Blood sample collection requires the assistance of multiple people, And special equipment is required, and the collection process is relatively time-consuming; ③Swine fever has a common antigen with bovine viral diarrhea virus and sheep border disease virus of the same genus, and the homology in gene structure reaches more than 60%

Analysis of the reasons for the above deficiencies in the prior art, (possibly) because: 1. the serological detection method is antigen and antibody reaction, and only once detection can not accurately judge the infection status and the detoxification status of pig herds, swine fever and bovine virus of the same genus There are 60% homologous in gene structure between CDV and ODV, and there is serological cross-reaction, which needs further verification

②Serological detection of antibodies, the level of detected antibodies cannot be quantitatively detected and analyzed, and serology can only evaluate the level of humoral immunity of the body; ③Serological evaluation of the health of pigs, evaluation of asymptomatic virus carriers and immune tolerance of pigs There are technical bottlenecks; ④ Serology needs to collect blood from the anterior vena cava, which is relatively laborious, time-consuming, and requires special equipment assistance.

However, due to the long operation time of the common PCR method currently used, it is easy to be polluted during the operation (compared to qPCR), which leads to certain limitations in the guiding significance of the diagnostic results for the prevention and control of epidemic diseases in pig farms.

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