Real-time fluorescent quantitative PCR detection primer probe set, kit and method of porcine circovirus type 3

Abstract

The invention belongs to the technical field of virus detection, and specifically relates to a real-time fluorescent quantitative PCR detection primer probe set, kit and method of porcine circovirus type 3. The detection accuracy is enhanced by designing primers on PCV3 cap protein in the invention for the first time; a fully closed tube operation is achieved in the detection method, the occurrence of false positive phenomenon is reduced, and at the same time, the high specificity of TaqMan probe technology and the high sensitivity of spectroscopy technology not only overcome the shortcoming of conventional PCR technology that can only achieve qualitative detection, but also solve the shortcoming of an SYBR Green method with poor specificity, and the detection method is more suitable for clinical testing; and the method has high sensitivity, good specificity and good reproducibility, the lowest detection limit is 11.8 copies / [mu]L, and the detection method is more sensitive than otherfluorescent quantitative PCR detection methods.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a primer probe set, a kit and a method for real-time fluorescent quantitative PCR detection of porcine circovirus type 3. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) was first isolated in 1974 from the pig kidney cell line PK-15 by German scholar Tischer et al. PCV is the smallest animal virus found so far. The diameter of the virus particle is about 17nm, and it is icosahedral. According to the sequence of discovery, pathogenicity and genome sequence, there are currently three types of circoviruses, namely PCV1 (Porcine circovirus 1), PCV2 (Porcine circovirus 2) and PCV3 (Porcine circovirus 3). PCV1 was first found to be non-pathogenic to pigs. The subsequently discovered PCV that can cause disease in pigs is called PCV2. PCV2 has been shown to be the main causative agent of diseases such as postweaning mμLtisystemic wasting ...

AI Technical Summary

Problems solved by technology

[0011] Another object of the present invention is to provide a real-time fluorescence quantitative PCR detection method of porcine circovirus type 3 TaqMan probe, the present invention for the first time by designing primers on the PCV3 cap protein to enhance the accuracy of detection; the detection method has achieved a fully closed tube At the same time, the high specificity of TaqMan probe technology and the high sensitivity of spectral technology not only overcome the shortcomings of conventional PCR technology that can only achieve qualitative detection, but also solve the problem of non-specificity of SYBR Green method. Strong and other shortcomings, more suitable for clinical detection; this method has high sensitivity, specificity and repeatability, and the lowest detection limit is 11.8 copies / μL, which is more sensitive than other fluorescent quantitative PCR detection methods

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