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Primer group for detecting clonorchis sinensis and detection method thereof

A clonorchis sinensis and detection method technology, applied in the field of parasite molecular detection, can solve the problems of low sensitivity, cumbersome detection method of clonorchis sinensis, long detection time, etc., and achieve low interference, high detection rate, and accuracy sex high effect

Inactive Publication Date: 2011-11-16
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to provide a detection primer set for Clonorchis sinensis in view of the defects of the prior art Clonorchis sinensis detection methods such as cumbersome, low sensitivity and long detection time

Method used

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  • Primer group for detecting clonorchis sinensis and detection method thereof
  • Primer group for detecting clonorchis sinensis and detection method thereof
  • Primer group for detecting clonorchis sinensis and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Detection of intermediate host C. sinensis

[0041] 1. Processing of intermediate host samples of Clonorchis sinensis to be tested:

[0042] 31 fish samples, 42 shrimp samples and 91 snail samples were collected in Yangshan County, Qingyuan City, and Sanshui District, Foshan City, Guangdong Province. Soak the collected snail samples in clear water for 24 hours and remove the shells, and remove the shells directly from the shrimp samples.

[0043] 2. Extraction of DNA from the intermediate host sample of Clonorchis sinensis to be tested:

[0044] (1) Add 200 μl proteinase K lysate to the treated sample to be tested, and digest it in a water bath at 55°C for 4-8 h;

[0045] (2) Take out the sample in step (1), add a mixture of phenol: chloroform: isoamyl alcohol with a volume ratio of 25:24:1, shake and mix at room temperature to form an emulsion, and then let it stand for 2-5 minutes;

[0046] (3) Centrifuge the mixture in step (2), and transfer the upper ...

Embodiment 2

[0059] Example 2 Determination of the temperature of the Clonorchis sinensis LAMP detection system

[0060] Use the total DNA of samples positive for Clonorchis sinensis as a template to determine the temperature of the LAMP detection system for the intermediate host of Clonorchis sinensis, the method is as follows:

[0061] 1. According to the method of Example 1, the total DNA of the intermediate host of Clonorchis sinensis was extracted.

[0062] 2. Carry out LAMP detection according to the method of Example 1, retain the total DNA of the intermediate host sample positive for Clonorchis sinensis, and use it as a template for determining the temperature of the LAMP detection system for the intermediate host of Clonorchis sinensis.

[0063] 3. Determination of the temperature of the LAMP detection system:

[0064] (1) Establish a loop-mediated isothermal amplification reaction system

[0065] LAMP reaction buffer (Reaction Buffer): Each 90ml LAMP reaction buffer contains ...

Embodiment 3

[0068] Example 3: Determination of the time of the LAMP detection system for the intermediate host of Clonorchis sinensis

[0069] Use the total DNA of samples positive for Clonorchis sinensis as a template to determine the time of the LAMP detection system for the intermediate host of Clonorchis sinensis, the method is as follows:

[0070] 1. According to the method of Example 1, the total DNA of the intermediate host of Clonorchis sinensis was extracted.

[0071]2. Carry out LAMP detection according to the method of Example 1, retain the total DNA of the intermediate host sample positive for Clonorchis sinensis, and use it as a template for determining the time of the Clonorchis sinensis intermediate host LAMP detection system.

[0072] 3. Determination of the time of the LAMP detection system:

[0073] 1) Establish a loop-mediated isothermal amplification reaction system

[0074] LAMP reaction buffer (Reaction Buffer): Each 90ml LAMP reaction buffer contains the followi...

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Abstract

The invention discloses a primer group for detecting clonorchis sinensis and a detection method thereof, belonging to the technical field of parasite molecular detection. The primer group for detecting clonorchis sinensis is as shown by SEQ ID NO: 1-6. According to the detection method, a sample to be detected is amplified through loop-mediated isothermal amplification reaction by the primers, the amplification product is detected, and whether the sample contains clonorchis sinensis can be determined by adding a fluorescent chromogenic reagent and observing with the naked eye in natural light, or by detecting whether positive gradient strips exist by an electrophoresis method. The primers for detection disclosed by the invention have high sensitivity and good specificity, and the detection method has high accuracy, simplicity of operation and broad application prospects.

Description

technical field [0001] The invention belongs to the technical field of parasite molecular detection, and relates to a primer and a detection method for detecting Clonorchis sinensis. Background technique [0002] Clonorchiasis caused by Clonorchis sinensis infection (also known as liver fluke) is mainly distributed in North Korea, South Korea, northern Vietnam, a small part of Russia and most of China. In China, clonorchiasis is mainly prevalent in Guangdong, Guangxi, Heilongjiang, Jilin, and Liaoning provinces. There are 27 provinces, municipalities, and autonomous regions with this disease epidemic or case reports, especially in Guangdong, Guangxi, Jilin, Heilongjiang, and Liaoning. (Fang Yueyi et al., 2008; Wei Dexiang et al., 1980; Wu Zhongbi et al., 1995; Zhao Weixian, 1994; Chen et al., 1984; Chen et al., 1991a; Chen et al.1991b; Kam et al., 1994; Woo et al ., 1998; Ko, 1991). [0003] The life cycle of Clonorchis sinensis is very complex, involving several intermedi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 伦照荣文婷温砚子陈月梁芳
Owner SUN YAT SEN UNIV
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