Institute of Environmental Science and Research, Christchurch Science Centre, New Zealand.
Appl Environ Microbiol. 2010 Mar;76(5):1533-44. doi: 10.1128/AEM.02215-09. Epub 2009 Dec 18.
To overcome some of the deficiencies with current molecular typing schema for Campylobacter spp., we developed a prototype PCR binary typing (P-BIT) approach. We investigated the distribution of 68 gene targets in 58 Campylobacter jejuni strains, one Campylobacter lari strain, and two Campylobacter coli strains for this purpose. Gene targets were selected on the basis of distribution in multiple genomes or plasmids, and known or putative status as an epidemicity factor. Strains were examined with Penner serotyping, pulsed-field gel electrophoresis (PFGE; using SmaI and KpnI enzymes), and multilocus sequence typing (MLST) approaches for comparison. P-BIT provided 100% typeability for strains and gave a diversity index of 98.5%, compared with 97.0% for SmaI PFGE, 99.4% for KpnI PFGE, 96.1% for MLST, and 92.8% for serotyping. Numerical analysis of the P-BIT data clearly distinguished strains of the three Campylobacter species examined and correlated somewhat with MLST clonal complex assignations and with previous classifications of "high" and "low" risk. We identified 18 gene targets that conferred the same level of discrimination as the 68 initially examined. We conclude that P-BIT is a useful approach for subtyping, offering advantages of speed, cost, and potential for strain risk ranking unavailable from current molecular typing schema for Campylobacter spp.
为了克服当前弯曲菌属分子分型方案的一些缺陷,我们开发了一种原型 PCR 二进制分型(P-BIT)方法。为此,我们研究了 58 株空肠弯曲菌、1 株拉里弯曲菌和 2 株大肠弯曲菌中 68 个基因靶标的分布情况。基因靶标是根据在多个基因组或质粒中的分布、作为流行因素的已知或假定状态选择的。为了比较,用彭纳血清分型、脉冲场凝胶电泳(SmaI 和 KpnI 酶)和多位点序列分型(MLST)方法对菌株进行了检测。P-BIT 对菌株的可分型率为 100%,多样性指数为 98.5%,而 SmaI PFGE 为 97.0%,KpnI PFGE 为 99.4%,MLST 为 96.1%,血清分型为 92.8%。对 P-BIT 数据的数值分析清楚地区分了三种弯曲菌属菌株,与 MLST 克隆复合体分类和以前的“高”和“低”风险分类有些相关。我们确定了 18 个基因靶标,它们具有与最初检测的 68 个基因靶标相同的区分能力。我们得出结论,P-BIT 是一种用于亚分型的有用方法,具有速度、成本和潜在的菌株风险分级优势,而这些优势是目前弯曲菌属分子分型方案所不具备的。