Double droplet digital PCR absolute quantitative detection Kit for European and American porcine reproductive and respiratory syndrome virus

Abstract

The invention discloses a double droplet digital PCR absolute quantitative detection kit for European and American porcine reproductive and respiratory syndrome virus. The primer probe combination comprises LV-F1R1P1 and VR2332-F1R1P1, wherein LV-F1R1P1 consists of LV-F1, LV-R1 and LV-P1, the LV-F1 is as shown in a sequence 1, LV-R1 is as shown in a sequence 2, and LV-P1 is as shown in a sequence 3; VR2332-F1R1P1 consists of VR2332-F1, VR2332-R1 and VR2332-P1, VR2332-F1 is as shown in a sequence 4, VR2332-R1 is as shown in a sequence 5, and VR2332-P1 is as shown in a sequence 6. The kit has an important application value on prevention and control of European and / or American porcine reproductive and respiratory syndrome viruses, and is favorable to epidemic situation control from the source and effective prevention of porcine epidemic large-scale outbreak.

Description

technical field [0001] The invention belongs to the field of virus detection, and in particular relates to a kit for absolute quantitative detection of porcine reproductive and respiratory syndrome virus European type and American type double droplet digital PCR. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). Characteristic highly contagious infectious disease. The disease broke out in the United States for the first time in 1987, and then appeared in Canada, Europe and Asia one after another. At present, it has become popular all over the world, causing huge economic losses every year. PRRS first appeared in my country in 1995, and then spread rapidly. In 2006, a pig "high fever disease" with high body temperature, high morbidity and high mortality broke out in my c...

AI Technical Summary

Problems solved by technology

Traditional PRRSV detection methods have disadvantages such as the need to use high-cost instruments and equipment, and the test takes a long time

Although RT-PCR and fluorescent RT-PCR have the characteristics of stronger specificity, high sensitivity, good repeatability, and high degree of automation compared with previous methods, the above methods can only achieve qualitative and semi-quantitative detection, and cannot detect viral nucleic acids. For accurate quantification, there are still some limitations in sensitivity and sensitivity specificity

[0004] The concept of digital PCR (Digital PCR, dPCR) was adopted and published by Bert Vogelstein as early as 1999, and its original intention was to obtain a large number of normal somatic cells from clinical samples (such as urine, lymph, plasma, stool, etc.) A small amount of mutant cells were detected in the sample, but because the consumables that could be used to dilute samples at that time were only 384-well plates, it could not reflect the core concept of digital PCR - "terminal dilution" (terminal dilution) very well.