Clonorchis sinensis real-time fluorescence polymerase chain reaction (PCR) detection kit and detection method

A clonorchis sinensis and detection kit technology, applied in the direction of biochemical equipment and methods, microbe measurement/inspection, etc., can solve the problems of cumbersome operation, low detection rate, time-consuming, etc., and achieve high detection accuracy and excellent results Accurate, save the effect of cumbersome operations

Active Publication Date: 2015-04-29
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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Problems solved by technology

Existing traditional detection methods mainly use morphological identificat

Method used

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  • Clonorchis sinensis real-time fluorescence polymerase chain reaction (PCR) detection kit and detection method
  • Clonorchis sinensis real-time fluorescence polymerase chain reaction (PCR) detection kit and detection method
  • Clonorchis sinensis real-time fluorescence polymerase chain reaction (PCR) detection kit and detection method

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Experimental program
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Embodiment 1

[0035] S10, take each 200 mg pretreated samples of Clonorchis sinensis, Paragonimus westermani, Toxoplasma gondii, Schistosoma japonicum, Angiostrongylus cantonensis, and Anisakis preserved in the Parasite Laboratory of Shenzhen Center for Disease Control and Prevention, and use Commercial tissue genomic DNA extraction kit, the specific extraction operation is carried out according to the instructions;

[0036] S20, performing PCR using the sample DNA extracted in step S10 as template DNA respectively, wherein the PCR system is as follows:

[0037]

[0038] Wherein, the above-mentioned PCR reaction reagents were all purchased from TaKaRa, and ABI-7500 fluorescent PCR instrument was used in the PCR process;

[0039] reaction process:

[0040]

[0041] Among them, in order to highlight the comparison of the amplification of each group of pathogenic bacteria, the positive group of the above-mentioned Clonorchis sinensis was used as two repeated controls; 2 O served as a n...

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Abstract

The invention provides a clonorchis sinensis real-time fluorescence polymerase chain reaction (PCR) detection kit which comprises a PCR specific primer and a fluorescent probe, wherein the PCR specific primer consists of a clonorchis sinensis ribosome 18S upstream primer having a base sequence shown as SEQ. ID. No. 1 in a sequence list and a clonorchis sinensis ribosome 18S downstream primer having a base sequence shown as SEQ. ID. No. 2 in the sequence list; the fluorescent probe has a base sequence shown as SEQ. ID. No. 3 in the sequence list; and the 5' end of the fluorescent probe is labeled with a fluorescent reporter group and the 3' end is labeled with a fluorescent quenching group. Compared with an existing detection method, the clonorchis sinensis real-time fluorescence PCR detection method disclosed by the invention can be used for reflecting the process of target gene amplification in real time and directly showing the copy number of the target group according to internal parameters or external parameters; the detection method has the advantages of being high in sensitivity, strong in specificity, short in cycle, high in flux and the like; meanwhile, the complex operation of gel electrophoresis is avoided by detecting a fluorescence signal by virtue of an optical system; and the detection method can be used for reducing the occurrence rate of false positive and potential hazard to human bodies and environment, and the detection method can be also used for shortening detection time.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a clonorchis sinensis real-time fluorescent PCR detection kit and a detection method. Background technique [0002] According to the monitoring results of foodborne parasitic diseases in parasite clinics in Shenzhen and the monitoring results of foodborne parasite projects in the food pollutant monitoring network in Shenzhen in recent years, the infection rate of clonorchis sinensis in aquatic products is gradually increasing. Therefore, it is necessary for us to detect food-borne parasites such as Clonorchis sinensis in aquatic products circulating on the market. The existing traditional detection methods mainly use morphological identification techniques, which are cumbersome, time-consuming, and low in detection rate. Contents of the invention [0003] The purpose of the embodiments of the present invention is to overcome the above-mentioned ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2561/113C12Q2563/107
Inventor 黄达娜张仁利吴春利阳帆李玥武伟华李晓恒高世同耿艺介
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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