Specific primers for loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis

A ring-mediated isothermal and angiostrongylus technology, applied in the field of thermal amplification detection, achieves the effect of simple and fast operation and short detection time

Inactive Publication Date: 2012-10-31
ZHEJIANG ACAD OF MEDICAL SCI +1
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Problems solved by technology

[0005] LAMP technology has been gradually used in medical testing since 2003. At present, it is mainly used in the detection of pathogenic microorganisms and the diagnosis of infectious diseases. Loop-mediated isothermal DNA amplification technology (LAMP) is used to quickly detect the 18s rRNA gene of Angiostrongylus cantonensis. There is no report in the literature at home and abroad

Method used

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  • Specific primers for loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis
  • Specific primers for loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis
  • Specific primers for loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis

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Embodiment

[0033] 1. Specific DNA sequence search:

[0034] The 18s rRNA gene sequence of Angiostrongylus cantonensis was retrieved from the American gene database-GENBANK, and the accession number is AY295804.1. LAMP primers were designed for this conserved target sequence. The specific target sequence is:

[0035] 1 attaagccat gcatgaggag ttcagcttta agtgaaactg cgaacggctc attagagcag

[0036]61 atgtgatta ttcggaaaat cctattggat aactgcggta attctggagc taatacatgc

[0037] 121 gtataaaccc tgactttcga aagggtgcaa ttattatagagc aaatcaatca ttttcggatg

[0038] 181 tagtttgctg actctgaata acgcagcata tcggcggctt gttcgccgat aatccgaaaa

[0039] 241 agtgtctgcc ctatcaacct gatggtagtc tattagtcta ccatggttat tacgggtaac

[0040] 301 ggagaataag ggttcgactc cggagaggga gccttagaaa cggctaccac atccaaggaa

[0041] 361 ggcagcaggc gcgaaactta tccaatcttg aatagatgag atagtgacta aaaataaaaa

[0042] 421 gaccattcct atggaacggt tatttcaatg agttgatcat aaaccttttt tcgagtatcc

[0043] 481 agtggagggc aagtctggtg ccagcagccg cggtaattcc...

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Abstract

The invention relates to the field of biotechnology, in particular to a method for rapidly detecting Angiostrongylus cantonensis by utilizing the loop-mediated isothermal amplification (LAMP) technology for the rapid detection technology of verminosis. In the method, two pairs of LAMP primers (FIP, BIP, F3 and B3) for detecting Angiostrongylus cantonensis are provided, and an LAMP method for detecting Angiostrongylus cantonensis by utilizing the two pairs of primers is provided. The method comprises preparation of a reaction template, utilization of an LAMP reaction system and an LAMP reaction program and judgment of detection results. The invention has the advantages of simple and convenient operation, low cost and easy observation of results, and is very suitable for on-site rapid detection of Angiostrongylus cantonensis.

Description

technical field [0001] The invention relates to the field of biological technology, in particular to a ring-mediated isothermal amplification detection method for Angiostrongylus cantonensis used in the rapid detection technology of parasitic diseases. Background technique [0002] Angiostrongylus cantonensis (Angiostrongylus cantonensis) was first discovered and named by the famous Professor Chen Xintao in the lung vessels of the Guangzhou house mouse. Snails and slugs are the main intermediate hosts. Humans are infected with Angiostrongylus cantonensis, also known as eosinophilic meningitis, by eating raw or half-cooked snail meat containing Angiostrongylus cantonensis larvae. Angiostrongylus cantonensis larvae mainly invade the central nervous system of the human body, manifested as meninges and encephalitis, meningitis and myelitis, which can cause death or disability. The main clinical symptoms are sharp headaches, which cannot even be subjected to any vibrations. The ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 陈睿陆绍红童群波张仪楼涤卓敏敏
Owner ZHEJIANG ACAD OF MEDICAL SCI
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