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Primer group and kit for detecting and identifying clonorchis sinensis and/or haplorchis taichui

A technology for Clonorchis sinensis and Mononorchis sinensis, which is applied in the field of primer sets and kits for detecting and identifying Clonorchis sinensis and/or Mononorchis sinensis, and can solve the problem that PCR detection primers and methods are not suitable for clinical diagnosis and There are problems such as deviation and low efficiency in disease monitoring work and identification test results, so as to reduce the cost of detection and identification, improve the efficiency of detection and identification, and reduce the false negative rate.

Inactive Publication Date: 2017-10-27
GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR DISEASE CONTROL & PREVENTION
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Problems solved by technology

At present, the two trematodes can be clearly identified by PCR based on the ribosomal DNA ITS region and / or COX1 (cytochrome oxidase subunit 1) gene of Clonorchis sinensis and the 5.8S rDNA of Monorches sinensis. The reason is that it is necessary to carry out PCR amplification reactions in two PCR reaction systems for different detection target genes; the detection process has the defects of cumbersome operation and low efficiency; due to the PCR amplification for different genes, there may be deviations in the identification detection results
It is generally believed that the current PCR detection primers and methods for the identification of Clonorchis sinensis and Monoorchis fanicum are not suitable for actual clinical diagnosis and disease monitoring

Method used

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  • Primer group and kit for detecting and identifying clonorchis sinensis and/or haplorchis taichui
  • Primer group and kit for detecting and identifying clonorchis sinensis and/or haplorchis taichui
  • Primer group and kit for detecting and identifying clonorchis sinensis and/or haplorchis taichui

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Embodiment 1

[0039]According to our long-term detection practice, for samples with low DNA template content (<1.0ng / μL), the sensitivity of PCR amplification using COX1 gene is significantly higher than that of PCR amplification based on ITS1 and ITS2 regions. This invention is based on the trematode genomic DNA extracted and purified by conventional methods from specimens such as digested adults, metacercariae and eggs, etc., and through a large number of comparative experiments in the early stage, the COX1 gene was selected as the PCR method to identify Clonorchis sinensis and fan. Target gene of Acanthorches sinensis. By optimizing the design of the primers, two pairs of highly specific and sensitive PCR detection primers were optimized; by optimizing the PCR reaction system and conditions, the simultaneous detection of the DNA of two trematodes in a single tube was successfully realized.

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Abstract

The invention relates to the technical field of biologics, and particularly relates to a primer group and a kit for detecting and identifying clonorchis sinensis and / or haplorchis taichui. The primer group is selected from a nucleotide sequence shown in SEQ ID NO.1 to SEQ ID NO.4. The primer group has the advantages of being good in specificity and high in amplification efficiency, primer dimer, non-specific amplification and cross reactions are avoided in the PCR (Polymerase Chain Reaction) amplification process, influence of non-target products to results is avoided, moreover, the amplification efficiency is high, and detection signals are intense.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set and a kit for detecting and identifying Clonorchis sinensis and / or Monorches sinensis. Background technique [0002] Clonorchis sinensis (Clonorchis sinensis) parasitizes the bile ducts in the human liver and can cause clonorchiasis and even cholangiocarcinoma. The occurrence and prevalence of Clonorchiasis sinensis are mainly related to the eating habits of people who prefer to eat raw freshwater fish and other foods. According to the survey, in Guangxi, where clonorchiasis is prevalent seriously, the average infection rate of Clonorchis sinensis among the population is 9.76%; among them, the population infection rate in individual endemic counties even exceeds 20%. Haplorchis taichui is a fluke of the family Heteromorphidae that lives in the intestinal tract. Monoorchis fanicum is commonly infected in people in Clonorchis endemic areas in Guangxi and other places. A...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143C12Q2547/101
Inventor 杨庆利
Owner GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR DISEASE CONTROL & PREVENTION
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