Duck virual hepatitis virus type I and duck plague virus dual-fluorescent quantitative PCR (polymerase chain reaction) method

Abstract

The invention provides a duck virual hepatitis virus type I and duck plague virus dual-fluorescent quantitative PCR (polymerase chain reaction) method. A pair of specific detection primers, namely DHV-I-F and DHV-I-R, and a fluorescent probe, namely DHV-I-R, are designed for duck virual hepatitis virus type I; a pair of specific detection primers, namely DPV-F and DPV-R, and a fluorescent probe, namely DPV-P, are designed for duck plague virus; and concentrations of the specific primers and the fluorescent probes of the dual-fluorescent quantitative PCR method for the duck virual hepatitis virus type I and the duck plague virus are determined. With the application of the dual-fluorescent quantitative PCR method provided by the invention, two viruses, namely the duck virual hepatitis virus type I and the duck plague virus, can be simultaneously detected and identified; the method has the advantages of being simple to operate, high in sensitivity, strong in specificity, good in repeatability and the like; the method not only can reduce cost but also can save precious time for epidemic surveillance and epidemic control; and in addition, effective means can be provided for researches on early rapid diagnosis and surveillance of of infectious duck diseases caused by the two viruses, epidemiological survey and the like.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a double fluorescence quantitative PCR method for type I duck hepatitis virus and duck plague virus. Background technique [0002] Duck viral hepatitis is an acute, severe, highly contagious and highly lethal viral infectious disease characterized by hepatitis. The pathogen is duck hepatitis virus (DHV), which mainly causes ducklings under 3 weeks of age to become ill. There are mainly three serotypes of duck hepatitis virus, namely type I, type II, and type III. These three serotypes are independent of each other. DHV-I is mainly prevalent in my country. Type Ⅰ duck hepatitis virus is most likely to be popular in spring from March to April. The mortality rate of ducklings under 3 weeks old is extremely high after infection. The mortality rate of ducklings under one week old can reach more than 80%. Reaching about 50%, has seriously jeopardized the healt...

AI Technical Summary

Problems solved by technology

Isolation culture has high requirements on culture environment, nutritional conditions, and separation operation, and it takes a long time; there are subjective factors in the interpretation of serological test results, the sensitivity and specificity are not high, and it is easy to cross with other microorganisms such as chlamydia Reaction and other problems; routine PCR detection has problems such as lack of specificity of primer binding, laboratory contamination, and presence of PCR inhibitors in clinical samples causing false negatives. After gene amplification, electrophoresis sequencing is required to verify, which is relatively time-consuming and costly. High; the real-time fluorescent probe PCR method distinguishes species based on sequence-specific probes, which improves the specificity and sensitivity of detection. For example, CN200810198528.9 discloses a molecular biology identification method for type I duck hepatitis virus, using fluorescent quantitative PCR method to identify type Ⅰ duck hepatitis virus, but this method can only detect type Ⅰ duck hepatitis virus, and cannot detect type Ⅰ duck hepatitis virus and duck plague virus at the same time for timely and accurate identification, which will easily delay epidemic monitoring and epidemic control

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