Ribosomal DNA ITS1 gene-based polymerase chain reaction (PCR) amplification kit for detecting clonorchis sinensis metacercaria and amplification primer

Abstract

The invention discloses a ribosomal DNA ITS1 gene-based polymerase chain reaction (PCR) amplification kit for detecting clonorchis sinensis metacercaria and an amplification primer. The kit comprises a PCR amplification reaction solution, wherein the PCR amplification reaction solution comprises 10 mmol / L of Tris-HCl, 50 mmol / L of KCl, 1.5 mmol / L of MgCl2, 0.8 mmol / L of dNTPs, 0.4 [mu]mol / L of positive primer, 0.4 [mu]mol / L of negative primer, 20 ng / [mu]L of DNA template, and 0.075 U / [mu]L of Taq enzyme. The kit disclosed by the invention is high in sensitivity and specificity, the detection is fast in speed and accurate, and an effective technical means is provided for making a diagnosis, giving treatment and preventing clonorchis sinensis infection.

Description

technical field [0001] The invention relates to the technical field of parasite detection, in particular to a PCR amplification kit and amplification primers for detecting Clonorchis sinensis metacercariae based on ribosomal DNAITS1 gene. Background technique [0002] Clonorchiasis (Clonorchiasis) is a food-borne zoonotic parasitic disease caused by infection with Clonorchis sinensis (Cs), which seriously endangers human health. The disease is mainly distributed in some countries in East Asia, Southeast Asia and Eastern Europe, such as China, South Korea, Laos and Vietnam. According to statistics, about 35 million people in the world are infected with this trematode, and 15 million people are distributed in various provinces and municipalities of my country (except Inner Mongolia, Qinghai, Tibet and Xinjiang), especially in Guangdong, Guangxi and the three northeast provinces. At present, there is no unified standard for the identification method of Clonorchis sinensis, and...

AI Technical Summary

Problems solved by technology

As we all know, microscopic examination is time-consuming, low-efficiency, and requires high clinical work experience for the examiner. Moreover, for the metacercariae that are closely related to Clonorchis sinensis, the eggs are similar in shape and cannot be distinguished by ordinary light microscopy. misjudgment

Immunodiagnostics, although more efficient than microscopy, are often limited by sensitivity and specificity

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