N-MDPV (novel Muscovy duck parvovirus) detection primer and probe and application thereof

Abstract

The invention provides a N-MDPV (novel Muscovy duck parvovirus) detection primer and probe and application thereof. The sequences of the primer and the probe are as follows: upstream primer NMM-F: 5'-TACGAATGAACAAACCAA-3'; downstream primer NMM-R: 5'-CGCTCTTAATATCTCCTCTA-3'; probe NMM-P: 5'-TGAACGAGCGAATGAGCCTTCC-3', wherein the 5'- end marks a fluorescence report group FAM, and the 3'- end marksa fluorescence quenching group Eclipse. The method has the advantages of high sensitivity, good stability, strong specificity and high repeatability and can be used for detecting N-MDPV infection, andlays a foundation for follow-up study of the pathogenesis of N-MDPV and development of molecular epidemiology.

Description

technical field [0001] The invention relates to a primer and probe for N-MDPV detection and application thereof, belonging to the field of duck diseases. Background technique [0002] Real-time fluorescent quantitative PCR method (Real-time PCR) is a method of detecting the total amount of product after each cycle of polymerase chain reaction (PCR) with fluorescent chemical substances in DNA amplification reaction. A method for quantitative analysis of a specific DNA sequence in a sample to be tested by using an internal or external reference method. Real-time fluorescent quantitative PCR is to detect the progress of PCR in real time through fluorescent signals during the PCR amplification process. 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...

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Problems solved by technology

[0007] The primers and probes of the real-time fluorescent quantitative PCR method for detecting N-GPV are designed for the VP3 gene (WangJ, et al. Development of a taqman-based real-time PCR assay for the rapid and specific detection of novel duck- origin goose parvovirus. Mol Cell Probes.2017, 34: 56-58. and Niu X, et al. Development of a TaqMan-based real-time PCRassay for the detection of Novel GPV. J Virol Methods. 2016, 237: 32-37. ), because N-MDPV is in the same genetic evolutionary branch as GPV on the VP3 gene, the above-mentioned TaqMan real-time fluorescent quantitative PCR method for detecting GPV may cause misjudgment and misdiagnosis due to potential viral gene recombination in the detection of N-MDPV (That is, the method can also detect N-MDPV)

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