Dual real-time fluorescent PCR detection reagents, kits and detection methods for Clonorchis sinensis and Angiostrongylus cantonensis

Abstract

The invention discloses a clonorchiasis sinensis and angiostrongyliasis cantonensis real-time fluorescence PCR (Polymerase Chain Reaction) detection reagent, and a kit and a detection method thereof, wherein the detection reagent comprises two groups of compositions taking conserved segments of clonorchiasis sinensis ITS1 gene and angiostrongyliasis cantonensis ITS2 gene as targets, and the two groups of compositions respectively comprise a pair of specific primers, a specific probe and an internal standard probe. The kit comprises PCR mixed solution, Taq DNA polymerase, DEPC-H2O, a positive quality control product, a negative quality control product and pseudovirus internal standard solution; the PCR mixed solution contains the detection reagent. The detection method comprises the steps of total DNA extraction, reaction composition preparation, amplification and result judgment. The reagent and the kit are strong in specificity, high in sensitivity, pollution-free, high-throughput, and capable of accurately detecting parasite larvae, invisible to the human eyes, in food, and the detection method is fast, specific and sensitive and capable of simultaneously detecting a great amount of samples.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a PCR detection reagent, a kit and a detection method thereof, in particular to a dual real-time fluorescent PCR detection reagent, a kit and a detection method of Clonorchis sinensis and Angiostrongylus cantonensis. Background technique [0002] Clonorchis sinensis (Clonorchis sinensis), also known as liver fluke, adult parasites in the bile duct of the liver, can cause clonorchiasis (clonorchiasis), also known as liver fluke disease. Clonorchis sinensis infection has been reported in China, Japan, Korea, Philippines, Singapore, Malaysia, Russia and other countries. In recent years, due to the diversification of food and the introduction of foreign eating habits, the infection rate of this disease in my country has gradually increased, and the infection rate of the population ranges from 1 to 30%. According to the first national survey of parasitic diseases, the national average infe...

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Problems solved by technology

[0004] The traditional detection of Clonorchis sinensis and Angiostrongylus cantonensis still remains in the stages of manual separation, microscopic examination, expert identification, etc., but the shortcomings of long detection cycle and extremely low sensitivity cannot be widely used

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