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Comparison of PCR Binary Typing (P-BIT), a New Approach to Epidemiological Subtyping of Campylobacter jejuni , with Serotyping, Pulsed-Field Gel Electrophoresis, and Multilocus Sequence Typing Methods
Author(s) -
Angela J. Cornelius,
Brent Gilpin,
Philip Carter,
Carolyn Nicol,
Stephen L. W. On
Publication year - 2010
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.02215-09
Subject(s) - multilocus sequence typing , pulsed field gel electrophoresis , subtyping , biology , typing , campylobacter jejuni , campylobacter , campylobacter coli , serotype , genetics , genotype , microbiology and biotechnology , gene , bacteria , computer science , programming language
To overcome some of the deficiencies with current molecular typing schema forCampylobacter spp., we developed a prototype PCR binary typing (P-BIT) approach. We investigated the distribution of 68 gene targets in 58Campylobacter jejuni strains, oneCampylobacter lari strain, and twoCampylobacter 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 threeCampylobacter 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 forCampylobacter spp.

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