GPV and N-GPV common type detection primer and probe

Abstract

The invention provides one group of GPV (Goose Parvovirus) and N-GPV (Novel-Goose Parvovirus) common type detection primer and probe. Sequences of the primer and the probe are as follows: an upstreamprimer NGG-F: 5'-TAGGGAGGAGTTAGAAGA-3'; a downstream primer NGG-R: 5'-CATCCATAGAATTGTCATAAGTA-3'; the sequence of a probe NGG-P is as follows: 5'-ACCTGGTAATTGTTCCTGCTTCTCT-3'; a 5'-end is labeled witha fluorescence reporter gene FAM and a 3'-end is labeled with a fluorescence quenching group Eclipse. A method has high sensitivity, good stability, strong specificity and good repeatability, can used for detecting N-GPV infection and can lay a foundation for subsequent researches of pathogenesis of the N-GPV and development of molecular epidemiology.

Description

Technical field [0001] The invention relates to a set of universal detection primers and probes for GPV and N-GPV, belonging to the field of poultry disease. Background technique [0002] Real-time fluorescent quantitative PCR (qPCR) is a method that uses fluorescent chemicals to detect the total amount of products after each polymerase chain reaction (PCR) cycle in a DNA amplification reaction. A method for quantitative analysis of specific DNA sequences in the sample to be tested by internal reference or external reference method. Real-time fluorescent quantitative PCR is the real-time detection of the PCR process through fluorescent signals during the PCR amplification process. Due to the exponential period of PCR amplification, the Ct value of the template has a linear relationship with the initial copy number of the template. The fluorescent probe method uses sequence-specific fluorescent labeled probes to detect products. The emergence of the probe method makes the specif...

AI Technical Summary

Problems solved by technology

At present, the primers and probes of the real-time fluorescent quantitative PCR method for detecting N-GPV are designed for the VP3 gene (Wang J, et al. Development of a taqman-based real-time PCR assay for therapeutic and specific detection of novel duck - origin goose parvovirus. Mol CellProbes. 2017, 34: 56-58. and Niu X, et al. Development of a TaqMan-based real-time PCR assay for the detection of Novel GPV. J Virol Methods. 2016, 237:32 -37.), but because N-MDPV has a long fragment (3116-4249) gene recombination between MDPV and GPV on the VP3 gene, the TaqMan probe qPCR detection method based on the VP3 gene for detecting N-GPV is less effective in detecting Due to the potential cross-reactivity of N-MDPV when using duck-derived disease materials, erroneous results may occur, which will affect the accurate diagnosis and treatment of the disease

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