Differentiation of Campylobacter jejuni and Campylobacter coli Using Multiplex-PCR and High Resolution Melt Curve Analysis

被引：13
|
作者：
Banowary, Banya [1 ,2 ,3 ]
Dang, Van Tuan [1 ,2 ,3 ]
Sarker, Subir [1 ,2 ,3 ]
Connolly, Joanne H. [1 ,2 ,3 ]
Chenu, Jeremy [4 ]
Groves, Peter [5 ]
Ayton, Michelle [1 ,2 ,3 ]
Raidal, Shane [1 ,2 ,3 ]
Devi, Aruna [6 ]
Vanniasinkam, Thiru [2 ,3 ,6 ]
Ghorashi, Seyed A. [1 ,2 ,3 ]
机构：
[1] Charles Sturt Univ, Sch Anim & Vet Sci, Wagga Wagga, NSW, Australia
[2] Graham Ctr Agr Innovat, NSW Dept Primary Ind, Wagga Wagga, NSW, Australia
[3] Charles Sturt Univ, Wagga Wagga, NSW, Australia
[4] Birling Avian Labs, Bringelly, NSW, Australia
[5] Univ Sydney, Sydney, NSW 2006, Australia
[6] Charles Sturt Univ, Sch Biomed Sci, Wagga Wagga, NSW, Australia
来源：
PLOS ONE | 2015年 / 10卷 / 09期
关键词：
FRAGMENT-LENGTH-POLYMORPHISM; SINGLE NUCLEOTIDE POLYMORPHISMS; FIELD GEL-ELECTROPHORESIS; REAL-TIME PCR; FECAL SAMPLES; GENE; IDENTIFICATION; ASSAY; STRAINS; REGION;
D O I：
10.1371/journal.pone.0138808
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
Campylobacter spp. are important causes of bacterial gastroenteritis in humans in developed countries. Among Campylobacter spp. Campylobacter jejuni (C. jejuni) and C. coli are the most common causes of human infection. In this study, a multiplex PCR (mPCR) and high resolution melt (HRM) curve analysis were optimized for simultaneous detection and differentiation of C. jejuni and C. coli isolates. A segment of the hippuricase gene (hipO) of C. jejuni and putative aspartokinase (asp) gene of C. coli were amplified from 26 Campylobacter isolates and amplicons were subjected to HRM curve analysis. The mPCR-HRM was able to differentiate between C. jejuni and C. coli species. All DNA amplicons generated by mPCR were sequenced. Analysis of the nucleotide sequences from each isolate revealed that the HRM curves were correlated with the nucleotide sequences of the amplicons. Minor variation in melting point temperatures of C. coli or C. jejuni isolates was also observed and enabled some intraspecies differentiation between C. coli and/or C. jejuni isolates. The potential of PCR-HRM curve analysis for the detection and speciation of Campylobacter in additional human clinical specimens and chicken swab samples was also confirmed. The sensitivity and specificity of the test were found to be 100% and 92%, respectively. The results indicated that mPCR followed by HRM curve analysis provides a rapid (8 hours) technique for differentiation between C. jejuni and C. coli isolates.
引用
收藏
页数：20
相关论文
共 50 条
  • [1] Evaluation of Two Multiplex PCR-High-Resolution Melt Curve Analysis Methods for Differentiation of Campylobacter jejuni and Campylobacter coli Intraspecies
    Banowary, Banya
    Van Tuan Dang
    Sarker, Subir
    Connolly, Joanne H.
    Chenu, Jeremy
    Groves, Peter
    Raidal, Shane
    Ghorashi, Seyed Ali
    [J]. AVIAN DISEASES, 2018, 62 (01) : 86 - 93
  • [2] Rapid Detection and Differentiation of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari in Food, Using Multiplex Real-Time PCR
    Mayr, A. M.
    Lick, S.
    Bauer, J.
    Thaerigen, D.
    Busch, U.
    Huber, I.
    [J]. JOURNAL OF FOOD PROTECTION, 2010, 73 (02) : 241 - 250
  • [3] Multiplex PCR assays for simultaneous identification of Campylobacter jejuni and Campylobacter coli
    Wieczorek, M
    Osek, J
    [J]. MEDYCYNA WETERYNARYJNA-VETERINARY MEDICINE-SCIENCE AND PRACTICE, 2005, 61 (07): : 797 - 799
  • [4] Detection and quantification of Campylobacter jejuni and Campylobacter coli using real-time multiplex PCR
    Toplak, N.
    Kovac, M.
    Piskernik, S.
    Mozina, S. Smole
    Jersek, B.
    [J]. JOURNAL OF APPLIED MICROBIOLOGY, 2012, 112 (04) : 752 - 764
  • [5] Differentiation of Campylobacter coli, Campylobacter jejuni, Campylobacter lari, and Campylobacter upsaliensis by a multiplex PCR developed from the nucleotide sequence of the lipid a gene lpxA
    Klena, JD
    Parker, CT
    Knibb, K
    Ibbitt, JC
    Devane, PML
    Horn, ST
    Miller, WG
    Konkel, ME
    [J]. JOURNAL OF CLINICAL MICROBIOLOGY, 2004, 42 (12) : 5549 - 5557
  • [6] A Comparison between Hippurate Hydrolysis and Multiplex PCR for Differentiating Campylobacter coli and Campylobacter jejuni
    Adzitey, Frederick
    Corry, Janet
    [J]. TROPICAL LIFE SCIENCES RESEARCH, 2011, 22 (01) : 91 - 98
  • [7] Identification of Campylobacter jejuni and Campylobacter coli from refrigerated chicken carcasses by Multiplex PCR
    Medeiros, Valeria de Mello
    Lopes Bricio, Silvia Maria
    Clementino, Maysa Mandetta
    [J]. VIGILANCIA SANITARIA EM DEBATE-SOCIEDADE CIENCIA & TECNOLOGIA, 2015, 3 (03): : 97 - 103
  • [8] Development of a multiplex PCR assay for identification of Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis
    Yamazaki-Matsune, Wataru
    Taguchi, Masurni
    Seto, Kazuko
    Kawahara, Ryuji
    Kawatsu, Kentaro
    Kumeda, Yuko
    Kitazato, Miyoshi
    Nukina, Masafumi
    Misawa, Naoaki
    Tsukamoto, Teizo
    [J]. JOURNAL OF MEDICAL MICROBIOLOGY, 2007, 56 (11) : 1467 - 1473
  • [9] Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing
    Mueller, Wolfgang
    Boehland, Corinna
    Methner, Ulrich
    [J]. RESEARCH IN VETERINARY SCIENCE, 2011, 91 (03) : E48 - E52
  • [10] Campylobacter jejuni and Campylobacter coli Genotyping by High-Resolution Melting Analysis of a flaA Fragment
    Merchant-Patel, Shreema
    Blackall, Patrick J.
    Templeton, Jillian
    Price, Erin P.
    Tong, Steven Y. C.
    Huygens, Flavia
    Giffard, Philip M.
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2010, 76 (02) : 493 - 499
12345