Dual real-time fluorescent RT-PCR identification and detection method of PRRSV classic strain and highly pathogenic mutant strain

Abstract

The invention discloses a dual real-time fluorescent RT-PCR method for identifying and detecting a PRRSV classic strain and a highly pathogenic mutant strain, and an RT-PCR cation primer and a probe against the PRRSV classic strain and the highly pathogenic mutant strain for the method. Compared with the prior art, the method is characterized by rapidness, simplicity, strong specificity, high sensitivity and good reliability and can simultaneously carry out large batch sample analysis, and one-time reaction can identify and diagnose whether a sample is infected by the PRRSV classic strain or infected by the PRRSV highly pathogenic mutant strain or infected by the two strains, thereby providing a powerful technical support for monitoring and controlling a PRRS epidemic disease and having good application prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for simultaneously detecting classical porcine reproductive and respiratory syndrome virus strains and highly pathogenic variant strains by using specific primers and probes. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). PRRSV was first isolated in the Netherlands and the United States, and served as the prototype strains of the European type (LV) and the American type (VR-2332), respectively. At present, PRRS has spread all over the countries and regions with developed pig industry in the world, and has brought huge economic losses to the world's pig industry and related industries. In recent years, PRRSV CH-1a, S1, HB-1 and HB-2 strains have been isolated successively in my country, all of which were identified as American PRRSV. Since June ...

AI Technical Summary

Problems solved by technology

[0003] At present, the commonly used methods for detecting PRRSV mainly include virus isolation and identification, serological detection and common RT-PCR, but these conventional methods are time-consuming, laborious, low-sensitivity and prone to false positives

Real-time fluorescent RT-PCR detection methods for all PRRSV have been established at home and abroad, but this method can only detect whether there is PRRSV infection, and cannot determine whether it is a classic PRRSV infection, or a highly pathogenic variant of PRRSV infection, or both. co-infection

In addition, the existing real-time fluorescent RT-PCR detection method for highly pathogenic variants of PRRSV can only detect highly pathogenic variants of PRRSV, and cannot determine whether it is a single highly pathogenic variant of PRRSV or a classic strain of PRRSV. Co-infection with highly pathogenic variants

Even if the two existing PRRSV real-time fluorescent RT-PCR methods are used in combination, it is still impossible to determine whether there is a co-infection problem, and cannot provide strong technical support for the rapid and effective monitoring of the PRRSV epidemic in my country

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