Clonorchis sinensis droplet type digital PCR detection kit and application thereof

A clonorchis sinensis and kit technology, which is applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as the inability to achieve absolute quantification and the undiscovered Clonorchis sinensis, and achieve good sensitivity and specific effects

Pending Publication Date: 2020-02-07
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The fluorescent quantitative PCR method relies on standards and standard curves to achieve quantitative detection of samples, but it is not yet possible to achieve absolute quantification
At present, there is no relevant report on the detection of Clonorchis sinensis using ddPCR method

Method used

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  • Clonorchis sinensis droplet type digital PCR detection kit and application thereof
  • Clonorchis sinensis droplet type digital PCR detection kit and application thereof
  • Clonorchis sinensis droplet type digital PCR detection kit and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0025] 1. Materials and methods

[0026] 1.1 Sample source and fecal genomic DNA extraction

[0027] 1.1.1 Sample source

[0028] Clonorchis sinensis-positive stool samples were obtained from Teng County, Guangxi, and tested positive by the modified Kato thick smear method. A total of 27 samples were obtained from the healthy control group. The stool samples of the healthy control group were obtained from Shanghai World Medical Center. The adults of Clonorchis sinensis and the eggs of Fasciola sinensis were provided by Xu Xuenian, a researcher at the Institute of Parasitic Diseases of the Chinese Center for Disease Control and Prevention; the adults of Hypoorchis sinensis were provided by Professor Wang Chunren of Heilongjiang Bayi Agricultural University; The DNA of the adult flukes of Cucumis brunii and Orthosimus sanpingi was provided by Zheng Bin, a researcher at the Institute of Parasitic Diseases Prevention and Control, Chinese Center for Disease Control and Prevention;...

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Abstract

The invention discloses a clonorchis sinensis droplet type digital PCR detection kit. The kit comprises a specific primer and a fluorescent probe, wherein the specific primer is designed according toa COI gene of a cytochrome C oxidase subunit 1 gene of clonorchis sinensis. In addition, the invention also discloses a primer and a probe for detecting clonorchis sinensis and an application of the primer and the probe in preparation of a clonorchis sinensis infection disease diagnosis product. An upstream primer of the primer is a sequence as shown in SEQ ID NO.1; the sequence of the downstreamprimer is shown as SEQ ID NO.2; and the probe has a sequence as shown in SEQ ID NO. 3. The invention successfully establishes the high-sensitivity detection kit capable of efficiently detecting clonorchis sinensis excrement DNA, and the detection kit is rapid, sensitive and specific and can be used for detecting clonorchis sinensis infection, especially mild infection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a PCR detection kit, in particular to a clonorchis sinensis droplet digital PCR detection kit. In addition, the present invention relates to the application of the Clonorchis sinensis droplet digital PCR detection kit. Background technique [0002] Clonorchis sinensis is an important food-borne disease. The latest survey shows that there are about 5.98 million infected people in my country. Clonorchis sinensis in the urban and suburban ecological areas of the Pearl River Delta, the western ecological areas of the Northeast Plain, and the Sanjiang Plain ecological area It was highly prevalent, and the infection rates were 23.36%, 9.44% and 8.53%. Dietary habit of eating raw fish is the main risk factor for Clonorchis sinensis infection in endemic areas. Clonorchiasis causes 275 370 DALYs lost, resulting in a severe economic burden. Acute and severe infections can cause non-specific ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6888C12Q1/6851C12Q2531/113C12Q2563/107
Inventor 沈玉娟徐梦曹胜魁张小凡曹建平
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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