Method for fast detecting trypanosome evansi by utilizing loop-mediated isothermal application technology

A ring-mediated isothermal and trypanosome technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of simple and fast operation and short detection time

Active Publication Date: 2012-11-28
ZHEJIANG ACAD OF MEDICAL SCI
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Problems solved by technology

[0005] LAMP technology has been gradually used in medical detection since 2003. At present, it is mainly used in the detection of pathogenic microorganisms and the diagnosis of infectious diseases. The loop-mediated isothermal DNA amplification technology (LAMP) is used to quickly detect the variation of Trypanosoma evans in various livestock Surface glycoprotein (VSG) gene has not been reported at home and abroad

Method used

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  • Method for fast detecting trypanosome evansi by utilizing loop-mediated isothermal application technology
  • Method for fast detecting trypanosome evansi by utilizing loop-mediated isothermal application technology
  • Method for fast detecting trypanosome evansi by utilizing loop-mediated isothermal application technology

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Embodiment

[0033] 1. Search for specific DNA sequence:

[0034] The VSG gene sequence of Trypanosoma evansi var. surface glycoprotein (VSG) was retrieved from the American gene database-GENBANK. The accession number is EF495337.1. LAMP primers were designed for the conservative target sequence. The specific target sequence is:

[0035] 1 catgcaaacc aaggcgctcg ttggcgtact cttatttgta ctgtatcgga gcacaacgga

[0036] 61 tgccgccaat gtagctctta aaggcaacgt ctggaagcca ttgtgcgaac tcgcggcagc

[0037] 121 gaccaggaac gggccaagcc acggcacggc gcacttcgca gcgatcgaaa atagcgtcga

[0038] 181 aacgtacact aagttaaaac taaagctctt gatttacgcg gcggccaaag gcagcaccac

[0039] 241 cgaagcaagc gcagcaagag ggttagcagc ggccgcagat agacacatac gagcagcggc

[0040] 301 caccacggcg aaagacaaaa gcagggtaat tctgcccgca gttgcctatg gcggcgaagt

[0041] 361 cgcaggggcg atttcatcgg cgctaaaatt tctaaagcac gcggttggca acagcaagtt

[0042] 421 ttgtgtgggc aaagccgacg gcacaaatgc cgacggtaac aacgaaatcg acgcgctagg

[0043] 481 gtgcggcgaa gccaactatg acacctcggc cccaggagac ag...

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Abstract

The invention relates to parasitic disease fast detection technology in the technical field of biology, and discloses a loop-mediated isothermal application (LAMP) method for detecting trypanosome evansi. In the method, two pairs of loop-mediated isothermal application primers FIP, BIP, F3 and B3 used for detecting the trypanosome evansi are adopted; and the LAMP method is used for detecting the trypanosome evansi by utilizing the two pairs of primers, and comprises the preparation of a reaction template, an LAMP reaction system, an LAMP reaction process and detection result judgment. The method has the advantages of simple and convenient operation, low cost, easily observable results and particular application to the field fast detection of the trypanosome evansi in livestock feeding farms and the like.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to a method for rapid detection of Trypanosoma evansi using loop-mediated isothermal amplification technology used in rapid detection of parasitic diseases. Background technique [0002] Trypanosoma evansi (Trypanosoma evansi) is a kind of hematomastigote parasites in the hematopoietic organs and blood of vertebrates. Horseflies and blood-sucking flies are the main transmission vectors, which can cause serious problems in domestic animals such as cattle, mules, horses, sheep and dogs. Trypanosomiasis evansi, also known as Sura disease, is mainly clinically manifested as high fever, anemia, jaundice and wormemia, and the acute mortality rate is very high. Trypanosoma evansi infection also leads to severe immunosuppression of the body, often leading to other opportunistic diseases, such as anthrax and Pasteurella infection. Saura disease is widespread in more than 20 provinces, cities, and au...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 陆绍红陈睿童群波楼涤卓敏敏
Owner ZHEJIANG ACAD OF MEDICAL SCI
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