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

A clonorchis sinensis and gene detection technology, applied in the field of PCR amplification kits and amplification primers, can solve problems such as low detection efficiency, limited sensitivity and specificity, and misjudgment, and achieve rapid detection, high sensitivity, and specificity strong effect

Inactive Publication Date: 2015-12-16
舟山出入境检验检疫局综合技术服务中心
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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 si...

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  • 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
  • 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
  • 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

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[0056] A PCR amplification kit for detecting Clonorchis sinensis metacercariae based on ribosomal DNA ITS2 gene, comprising a PCR amplification reaction liquid, said PCR amplification reaction liquid comprising: Tris-HCl10mmol / L with pH8.3, KCl50mmol / L , MgCl 2 1.5mmol / L, dNTPs0.8mmol / L, forward primer 0.4μmol / L, reverse primer 0.4μmol / L, DNA template 20ng / μL, Taq enzyme 0.075U / μL. The sequence of the forward primer is: 5'-ATAAACTATCACGACGCCCAA-3' (SEQ ID NO.1), and the sequence of the reverse primer is: 5'-GGTGCAAAACAGATTTGCATC-3' (SEQ ID NO.2).

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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 ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 李孝军陈璐敏白颉单长林沈飚杨赛军杜爱芳王素华
Owner 舟山出入境检验检疫局综合技术服务中心
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