Same-Day Subtyping of Campylobacter jejuni and C. coli Isolates by Use of Multiplex Ligation-Dependent Probe Amplification–Binary Typing

Abstract

e Campylobacteriosis is the most commonly reported form of human bacterial gastroenteritis in the world. Sound identification of infectious sources requires subtyping, but the most widely used methods have turnaround times measured in days and require specialist equipment and skills. A multiplex ligation-dependent probe amplification-binary typing (MBiT) assay was developed for subtyping Campylobacter jejuni and Campylobacter coli. It was tested on 245 isolates, including recent isolates from Belgium and New Zealand, and compared to multilocus sequence typing (MLST). When used in an outbreak setting, MBiT identified the predominant genotype and possible additional cases days before pulsed-field gel electrophoresis (PFGE) results were available. MBiT was more discriminatory than MLST and, being a single assay with results produced within 6 h, was more rapid and cost-effective than both MLST and PFGE. In addition, MBiT requires only basic molecular biology equipment and skills.

Comparative genomic fingerprinting of Campylobacter: application in routine public health surveillance and epidemiological investigations

“…Protocols could be designed to target public health investigation following the implementation of routine CGF40 subtyping, similar to those in place through PulseNet Canada for Salmonella, Escherichia coli, and Listeria [28,29]. Our data support other findings that small outbreaks are relatively common [30], beyond those detected through routine public health follow up. Recent work has shown the potential for WGS in the analysis of Campylobacter isolates in an epidemiological context [31][32][33] and we anticipate that WGS will play an increasingly prominent role in epidemiological investigations.…”

Concentrations of Campylobacter spp., Escherichia coli, Enterococci, and Yersinia spp. in the Feces of Farmed Red Deer in New Zealand

“…Colonies identified as Campylobacter jejuni and Campylobacter coli were subtyped using a multiplex ligation‐dependent probe amplification‐binary typing (MBiT) assay (Cornelius et al, 2014). In brief, isolates for MBiT analysis were purified to obtain single colonies, and one colony was then resuspended in 250 μL of 2% Chelex in H 2 O.…”

Inquiring into the Gaps of Campylobacter Surveillance Methods

“…The highly variable genome of these Campylobacter species has raised concerns regarding the application of typing methodologies based on DNA sequence information such as the MLST, flaA SVR, and the MOMP [ 111 , 112 ]. This obstacle paved the way towards combining methodologies for a more accurate Campylobacter populations investigation, such as Multiplex Ligation-Dependent Probe Amplification–Binary Typing [ 102 , 113 ]. Genomic mosaicism can adversely impact the understanding of closely related species and the use of few genetic loci can lead to erroneous results.…”