PCR (Polymerase Chain Reaction) primer for simultaneously detecting RHDVa (Rabbit Hemorrhagic Disease Virus a) and RHDVb (Rabbit Hemorrhagic Disease Virus b) as well as kit and non-diagnosis-purpose detection method thereof

Abstract

The invention relates to the technical field of detection and in particular relates to a PCR (Polymerase Chain Reaction) primer for simultaneously detecting RHDVa (Rabbit Hemorrhagic Disease Virus a)and RHDVb (Rabbit Hemorrhagic Disease Virus b) as well as a kit and a non-diagnosis-purpose detection method thereof. The PCR primer is composed of a rabbit hemorrhagic disease virus mutation type a detection primer and a rabbit hemorrhagic disease virus mutation type b detection primer; the rabbit hemorrhagic disease virus mutation type a detection primer is composed of an upstream primer shown as SEQ ID NO: 1 and a downstream primer shown as SEQ ID NO: 2; the rabbit hemorrhagic disease virus mutation type b detection primer is composed of an upstream primer shown as SEQ ID NO: 3 and a downstream primer shown as SEQ ID NO: 4. The PCR primer provided by the invention has the advantages of good specificity, sensitivity and accuracy, no need to large-size instruments and equipment or subsequent sequencing verification, cost reducing, convenience for operation, rapidness and the like.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a PCR primer for simultaneous detection of RHDVa and RHDVb, a kit thereof and a non-diagnostic detection method. Background technique [0002] The production and trade of rabbit products is an important part of the livestock industry. my country's domestic rabbit slaughter, stock and rabbit meat production account for 40%, 25% and 41.09% of the world's total respectively. Rabbit hemorrhagic disease (commonly known as rabbit plague) is a common disease that threatens the life and health of rabbits, and it has a serious impact on the production of rabbit products. Under natural conditions, the disease mainly affects adult rabbits over 2 months old. It is characterized by respiratory hemorrhage, parenchymal organ congestion, enlargement, hemorrhage, and liver necrosis. It is a fulminant epidemic, with short incubation period, acute onset, short course of disease (rabbits usuall...

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Problems solved by technology

In addition, the specificity of the PCR reaction mainly depends on the specific combination of primers and target sequences. Due to the genetic variation characteristics of the virus itself, the above methods face the risk of false negative or false positive.

For the test results of PCR, if further confirmation is required, the nucleic acid sequence of the product needs to be determined, resulting in an increase in the test cycle and cost; although fluorescent quantitative PCR can improve the reliability of the test by using specific probes or comparing PCR amplification curves However, the result still depends on the specific combination of the primer and the target sequence, and the risk of false negative or false positive results cannot be avoided, and the method is complicated to operate, requires special equipment, and the cost of reagents is expensive

Therefore, the above methods cannot adapt to the development trend of international trade and meet the current and future needs of rapid and accurate inspection of various types of rabbit hemorrhagic disease virus quarantine in China.

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