Development and Optimization of a Loop-mediated Isothermal Amplification Assay for Detection of Campylobacter jejuni

doi:10.3969/j.issn.1000-7091.2013.03.040

Abstract

Abstract: In this study， the specific mapA gene was used to design a set of primers to develop a loop-mediated isothermal amplification ( LAMP) assay for detection of C． jejuni． To optimize the LAMP assay， the reaction components in the assay were screened to determine the optimal concentration． The final LAMP assay comprised 1．6μmol /L each of inner primers FIP and BIP，0．2 μmol /L each of outer primers F3 and B3,8 mmol /L Mg2 + ，1．4mmol /L dNTP，0 ．8 mol /L Betaine． The specificity test showed that the assay correctly identified all C． jejuni strains but not 13 other bacterial species． The sensitivity of the assay was 100 fg per test tube for C． jejuni genomic DNA and 7． 5 cfu per test tube for C． jejuni bacterial culture． This LAMP assay is a rapid and simple tool for detection of C． jejuni and will provide an essential role in facilitating early diagnosis of this organism from animal products．

Key words: Campylobacter jejuni, Loop-mediated isothermal amplification assay, Detection, Optimization

CLC Number:

S855.1

Xu Zijian, Yang Bing, Su Xia, Zhou Hongzhuan, Shi Aihua, Xu Fuzhou. Development and Optimization of a Loop-mediated Isothermal Amplification Assay for Detection of Campylobacter jejuni[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(3): 222-226. doi: 10.3969/j.issn.1000-7091.2013.03.040.

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http://www.hbnxb.net/EN/Y2013/V28/I3/222

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