Primer and probe for detecting echinococcus multilocularis in canine feces, and kit of primer and probe

A technology of Echinococcus multilocularis and a kit is applied in the field of genetic engineering, which can solve the problems of labor-consuming, time-consuming, complicated process and the like, and achieve the effects of simple operation, accurate results and controllable cost.

Inactive Publication Date: 2017-11-21
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The object of the present invention is to provide a primer and a probe for detecting Echinococcus multilocularis in dog feces and a kit thereof, said primer and probe for detecting Echinococcus multilocularis in dog feces The method and the kit thereof shall solve the technical problems of complex, manpower-consuming and time-consuming process of detecting Echinococcus multilocularis in dog feces in the prior art

Method used

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  • Primer and probe for detecting echinococcus multilocularis in canine feces, and kit of primer and probe
  • Primer and probe for detecting echinococcus multilocularis in canine feces, and kit of primer and probe
  • Primer and probe for detecting echinococcus multilocularis in canine feces, and kit of primer and probe

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Acquisition of real-time PCR primer-specific primers and probes for differentiating Echinococcus multilocularis in dog feces.

[0023] According to the mitochondrial cytochrome c oxidase subunit I gene cox1 of Echinococcus multilocularis (GenBank TM accession number: AB813188), using Primer Express 3.0 software to design and synthesize Taqman fluorescent quantitative PCR specific primers and probes, including:

[0024] A group of real-time fluorescent quantitative PCR primers and probes for specifically amplifying Echinococcus multilocularis, its nucleotide sequence is:

[0025] Upstream primer EfF: 5'-GATTCCTTCTTTAGTTTTGTTGTTGATTAG-3',

[0026] Downstream primer EfR: 5'-CGAACTAAAAACTCTAGAAACACCTGCT-3',

[0027] Probe EF: 5'-F-TGGTTGGACTTTTTATCCTCCATTGTCTTTCTTCA-Q-3'.

[0028] Wherein, F is the fluorescent reporting group FAM, and Q is the fluorescent quenching group TAMRA.

[0029] The amplification system used in the real-time quantitative PCR reaction is 25 μL, a...

Embodiment 2

[0036] Example 2 Detection application of Echinococcus multilocularis in dog feces

[0037] According to the method of Example 1, 30 dog feces samples from Ganzi Prefecture, Sichuan Province were tested by fluorescent quantitative PCR, and then the 30 samples were sequenced and tested using the gold standard of etiology and genetic testing - gene sequencing. The results showed that the results of Taqman fluorescent quantitative PCR experiment were 100% consistent with the results of etiology and gene sequencing.

[0038]

[0039]

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Abstract

The invention provides a primer for detecting echinococcus multilocularis in canine feces, wherein the sequence of an upstream primer is as shown in SEQ ID No. 1; and the sequence of a downstream primer is as shown in SEQ ID No. 2. The invention also provides a probe for detecting the echinococcus multilocularis in the canine feces, wherein the sequence is as shown in SEQ ID No. 3. The invention further provides a kit for detecting the echinococcus multilocularis in the canine feces; the kit comprises the primer for detecting the echinococcus multilocularis in the canine feces and the probe for detecting the echinococcus multilocularis in the canine feces, wherein the sequence of the upstream primer is as shown in SEQ ID No. 1; the sequence of the downstream primer is as shown in SEQ ID No. 2; and the sequence of the probe is as shown in SEQ ID No. 3. In the invention, the fluorescence quantitative PCR reaction of the echinococcus multilocularis can be effectively amplified by utilizing the primer and the probe, and the echinococcus multilocularis infection can be specifically identified and diagnosed.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to a method for detecting parasites, in particular to a primer, a probe and a kit for detecting Echinococcus multilocularis in dog feces. Background technique [0002] The morphology and life cycle of Echinococcus multilocularis are similar to those of Echinococcus multilocularis, but the adults mainly parasitize foxes, the intermediate host is rodents or insectivores, and the larval stage is Echinococcus multilocularis (also known as alveolar larvae). It causes severe alveolar hydatid disease or multilocular hydatid disease in humans. The distribution area of ​​Echinococcus multilocularis is more limited than that of Echinococcus granulosus, and it is mainly prevalent in the high latitudes of the northern hemisphere, from northern Canada, Alaska, to Hokkaido, Japan, and Siberia, Russia. Throughout the cold regions and tundra areas of North America, Europe and Asia. [0003] The a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2561/101C12Q2563/107C12Q2561/113
Inventor 陈家旭肖宁陈韶红艾琳陈木新田利光储言红卢艳蔡玉春吴秀萍俞英昉宋鹏陈海宁李浩
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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