LAMP (loop-mediated isothermal amplification) kit for rapid detection of duck plague viruses

Abstract

The invention discloses an LAMP (loop-mediated isothermal amplification) kit for rapid detection of duck plague viruses. According to the invention, a set of complete LAMP system is constructed; an LAMP amplification method can be achieved with the application of a water-bath kettle or a heat-preserving cup; results are visible to naked eyes; and the LAMP amplification method is relatively simple to operate and high in sensitivity, and the method is specially suitable for on-site quarantine at a basic level. A detection duration (including a sample processing time) is shortened from 3-4h to 2h, detection limit reaches 1pg / mL and specificity is 100%, without occurrence of false positive and false negative. With the application of the kit and a DPV pathogen PCR detection method (GB / T 22332-2008), duck plague viruses in 200 sick duck samples can be detected, and a coincidence rate of the two methods reaches 100%. The kit provided by the invention has the characteristics of being efficient, strong in specificity, high in sensitivity and the like, and a requirement of rapidly detecting samples in a large batch can be satisfied. A subsequently established LAMP diagnosis formula is applicable to on-site rapid detection and is accurate in result and convenient to use.

Description

technical field [0001] The invention relates to the fields of livestock breeding and disease prevention and control, in particular to a rapid detection kit for duck plague virus LAMP. Background technique [0002] Duck plague (Duck plague, DP), also known as duck viral enteritis (Duck viral enteritis, DVE), is a disease of waterfowl such as ducks, geese and swans caused by duck plague virus (Duck plague virus, DPV) of the Herpesviridae family. An acute, febrile, septic infectious disease, the main clinical features are elevated body temperature, numbness of the legs, diarrhea, tearing, and swelling of the head and neck of some sick ducks, so the disease is commonly known as "big head plague" in some parts of my country. Duck plague was first reported in the Netherlands in 1923, and then the occurrence and prevalence of the disease were reported in many duck-raising countries around the world. my country first discovered the disease in Guangzhou in 1957. Over the years, duck ...

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Problems solved by technology

However, the virus isolation and identification method may not cause clinical symptoms in the case of low-virulence or non-pathogenic strains; although the result of serological neutralization test is accurate, the procedure is complicated, time-consuming and laborious, and cannot be used for rapid diagnosis and large-scale samples detection; ELISA is currently the most widely used method, which is economical, simple, and fast, but its sensitivity is slightly lower than that of PCR technology; traditional PCR technology has the characteristics of high sensitivity and strong specificity, but this detection The cost of the method is high, the requirements for operator skills and reaction instruments are high, and the amplification reaction time is long, which is not conducive to rapid detection and popularization and application in grassroots laboratories

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