PCR (polymerase chain reaction) amplification kit for detecting metacercaria of Cs (clonorchis sinensis) on basis of ribosomal DNA (deoxyribose nucleic acid) ITS2 (internal transcribed spacer 2) and amplification primer

Abstract

The invention discloses a PCR (polymerase chain reaction) amplification kit for detecting metacercaria of Cs (clonorchis sinensis) on the basis of ribosomal DNA (deoxyribose nucleic acid) ITS2 (internal transcribed spacer 2) and an amplification primer. The PCR amplification kit comprises a PCR amplification reaction liquid, wherein the PCR amplification reaction liquid comprises 10 mmol / L of Tris-HCl , 50 mmol / L of KCl, 1.5 mmol / L of MgCl2, 0.8 mmol / L of dNTPs (deoxyribonucleoside-5'-triphosphate), 0.4 mu mol / L of a forward primer, 0.4 mu mol / L of a reverse primer, 20 ng / mu L of a DNA template and 0.075 U / mu L of a Taq enzyme. The PCR amplification kit is high in sensitivity and high in specificity, can detect metacercaria of Cs quickly and accurately and provides an effective technical means for diagnosis and prevention of Cs 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 DNITS2 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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