Primer and probe for real-time fluorescence quantitative PCR detection of two genotypes of duck circovirus

Abstract

The invention relates to a primer and probe for real-time fluorescence quantitative PCR detection of two genotypes of duck circovirus. Sequences of the primer and the probe are as follows: an upstreamprimer DuCV-F: 5'-CAGTTTGTKGCTAARACVTTG-3'; a downstream primer DuCV-R: 5'-AGTTTATTGGRAASGGGAGG-3'; a probe DuCV-1-pobe: 5'-AGAGCGAGTTTGACCTYT-3'; a probe DuCV-2-pobe: 5'-TTTGATTTGTCCGCCTTAT-3', wherein the 5'- end of the DuCV-1-pobe is labelled with a fluorescent reporter group Cy5; and the 5'- end of the DuCV-2-pobe is labelled with a fluorescent reporter group FAM. The method is high in sensitivity and specificity, and can be used for genotype identification of two kinds of duck circovirus.

Description

technical field [0001] The invention belongs to the field of poultry disease, and in particular relates to a primer and a probe for real-time fluorescent quantitative PCR detection of two genotypes of duck circovirus. Background technique [0002] Real-time fluorescent quantitative PCR method (Real-time PCR) is a method of detecting the total amount of products after each cycle of polymerase chain reaction (PCR) with a fluorescent chemical substance in a DNA amplification reaction. During the PCR amplification process, real-time fluorescent quantitative PCR can detect the progress of PCR in real time through fluorescent signals. Due to the exponential phase of PCR amplification, there is a linear relationship between the Ct value of a template and the initial copy number of the template. The fluorescent probe method uses sequence-specific fluorescently labeled probes to detect products. The emergence of the probe method greatly improves the specificity of quantitative PCR t...

AI Technical Summary

Problems solved by technology

In 2018, Korean scholars studied the pathogenicity of D11-JW-008 strain (belonging to DuCV-1 branch) to Peking duck (24d) and found that apoptosis could be detected in the bursa of Peking duck after infection, but the experimental group and Similar to the control group, typical clinical symptoms of DuCV infection (such as emaciation, disheveled feathers, and weight loss, etc.) were not observed

At present, there are no research reports on real-time fluorescent quantitative primer sets and probe sets that can specifically diagnose DuCV-1 and DuCV-2 infections. This method can not only be used for epidemiological detection (reaching the effect of conventional PCR detection, But the sensitivity is higher), and it can accurately quantify the specific subtype infection of DuCV-1 and DuCV-2, and at the same time accurately distinguish different infection types

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