Primer sets and kits for detection and identification of Clonorchis sinensis and/or Monorches sinensis
A technique 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 Disease monitoring work, discriminating test results, low efficiency and other problems, to achieve the effect of reducing the cost of detection and identification, low false negative rate, and high sensitivity
Inactive Publication Date: 2020-01-14
GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR DISEASE CONTROL & PREVENTION
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Problems solved by technology
Currently, methods based on the Clonorchis sinensis ribosomal DNA ITS region and / or COX 1 (cytochrome c oxidase subunit 1) gene and the PCR method of the 5.8S rDNA of Monorches sinensis can clearly identify these two kinds of trematodes, but the defect of this technique is that it needs to target different detection genes in two PCR reaction systems Carry out PCR amplification reaction; the detection process has the defects of cumbersome operation and low efficiency; due to PCR amplification for different genes, there may be deviations in the identification and 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
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Embodiment 1
[0039] According to our long-term detection practice, for samples with low DNA template content ( COX The sensitivity of PCR amplification of 1 gene was 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. COX1 gene was used as the target gene for PCR method to identify Clonorchis sinensis and Monorches 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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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 infection rate of population in individual endemic counties even exceeds 20%. Monoorchis fanicum ( 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 area...
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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143C12Q2547/101
Inventor 杨庆利
Owner GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR DISEASE CONTROL & PREVENTION