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Kit for assaying echinococcus in lesion tissue or dog feces by multiplex recombinase-aid amplification (RAA) and multiple PCR as well as assay method

A technology of Echinococcus multilocularis and Echinococcus, which is applied in the fields of agriculture and animal quarantine, can solve the problems of high cost, time-consuming and laborious, etc., and achieve the effects of saving detection costs, avoiding high false positives, and improving amplification performance

Active Publication Date: 2020-01-07
中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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Problems solved by technology

However, the detection of a single insect species is expensive, time-consuming and laborious

Method used

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  • Kit for assaying echinococcus in lesion tissue or dog feces by multiplex recombinase-aid amplification (RAA) and multiple PCR as well as assay method
  • Kit for assaying echinococcus in lesion tissue or dog feces by multiplex recombinase-aid amplification (RAA) and multiple PCR as well as assay method
  • Kit for assaying echinococcus in lesion tissue or dog feces by multiplex recombinase-aid amplification (RAA) and multiple PCR as well as assay method

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Effect test

Embodiment 1

[0060] 1. Materials and methods

[0061] 1. Materials

[0062] 1.1 sample

[0063] 1.1.1 Echinococcus granulosus G1 and Echinococcus multilocularis tissue samples were provided by the Health Education Center and Key Laboratory of the Institute of Parasitic Diseases Prevention and Control, Chinese Center for Disease Control and Prevention of the applicant, and stored at -20°C.

[0064] 1.1.2 Collection of samples: Infected tissue samples were collected from Qinghai, Sichuan, Xinjiang, and laboratory-infected animal tissue samples (19), and negative dog feces samples were collected from Shanghai.

[0065] 1.2 Reagents

[0066] Blood / tissue DNA extraction kit and stool DNA extraction kit were purchased from Qiagen, USA, Taq DNA polymerase (5U / μL), dNTPMix, Mgcl 2 , 10×Reaction Buffer was purchased from Shanghai Takara Company, and PCR primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd. 50×TAE buffer and Agarose agarose were purchased from Beijing Suo Laibao Te...

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Abstract

The invention discloses a kit for assaying echinococcus in lesion tissue or dog feces by multiplex recombinase-aid amplification (RAA) and multiple PCR. Meanwhile, the kit is also used for assaying echinococcus granulosus G1 and echinococcus multilocularis. The kit comprises primers as follows: echinococcus granulosus specific primers shown in SEQ ID NO.1 and SEQ ID NO.2 and echinococcus multilocularis specific primers shown in SEQ ID NO.3 and SEQ ID NO.4. In addition, the invention further discloses a multiplex RAA and multiplex PCR assay method for the echinococcus granulosus G1 and echinococcus multilocularis. Various verifications prove that the kit has characteristics of rapidness, sensitivity and high specificity, complex steps for repeated assay are eliminated, two genes can be assayed simultaneously in one reaction, functions of pathogens are made clear, the workload is reduced greatly, the assay time is shortened, the assay cost is reduced, and the kit has a greater development trend.

Description

technical field [0001] The invention relates to the technical fields of agriculture and animal quarantine, in particular to a detection method for Echinococcus granulosus G1 and Echinococcus multilocularis, and in particular to a multiple recombination enzyme for Echinococcus granulosus G1 and Echinococcus multilocularis Mediated amplification technology (Recombinase-aid Amplification, RAA) and multiplex PCR detection methods and kits. Furthermore, the present invention relates to primers for the simultaneous detection of Echinococcus granulosus G1 and Echinococcus multilocularis. Background technique [0002] Echinococciosis, also known as hydatidosis, is a chronic parasitic disease caused by the larvae of Echinococcus tapeworm parasitizing human tissues and organs, and it is also an important zoonotic disease. Echinococcus has a complex life cycle, a wide range of epidemics and many hosts, and the intermediate hosts and final hosts are also different in different countrie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12Q1/6844C12N15/11
CPCC12Q1/6888C12Q1/686C12Q1/6844C12Q2600/16C12Q2537/143C12Q2521/507Y02A50/30
Inventor 周鸿让肖宁余晴陈木新王莹艾琳王晓玲许秋利
Owner 中国疾病预防控制中心寄生虫病预防控制所国家热带病研究中心
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