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Loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis

A loop-mediated isothermal, angiostrongylus technology, applied in the field of temperature amplification detection, achieves the effect of simple and rapid operation and short detection time

Inactive Publication Date: 2010-12-01
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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  • Loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis
  • Loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis
  • Loop-mediated isothermal amplification (LAMP) detection method of Angiostrongylus cantonensis

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Embodiment

[0033] 1. Search for specific DNA sequence:

[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 the conservative target sequence. The specific target sequence is:

[0035] 1 attaagccat gcatgaggag ttcagcttta agtgaaactg cgaacggctc attagagcag

[0036] 61 atgtgattta ttcggaaaat cctattggat aactgcggta attctggagc taatacatgc

[0037] 121 gtataaaccc tgactttcga aagggtgcaa ttattagagc 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 agctccacta gt...

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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 biotechnology, in particular to an Angiostrongylus cantonensis ring-mediated isothermal amplification detection method used in the rapid detection technology of parasitic diseases. Background technique [0002] Angiostrongylus cantonensis (Angiostrongylus cantonensis) was first discovered and named in the pulmonary blood vessels of Guangzhou domestic mice by the famous Chinese Professor Chen Xintao. Snails and slugs are the main intermediate hosts. People are infected with Angiostrongylus cantonensis by eating raw or half-lived snail meat containing Angiostrongylus cantonensis larvae, also known as eosinophilic meningitis. Angiostrongylus cantonensis larvae mainly invade the human central nervous system, manifesting as meningitis and encephalitis, meningitis and myelitis, which can cause death or disability. The main clinical symptoms are sharp headaches, which can not even be subjected to any vibrations. The headaches...

Claims

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Application Information

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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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