A pan-European ring trial to validate an International Standard for detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus in seafoods

Abstract

Globally, vibrios represent an important and well-established group of bacterial foodborne pathogens. The European Commission (EC) mandated the Comite de European Normalisation (CEN) to undertake work to provide validation data for 15 methods in microbiology to support EC legislation. As part of this mandated work programme, merging of ISO/TS 21872-1:2007, which specifies a horizontal method for the detection of V. parahaemolyticus and V. cholerae, and ISO/TS 21872-2:2007, a similar horizontal method for the detection of potentially pathogenic vibrios other than V. cholerae and V. parahaemolyticus was proposed. Both parts of ISO/TS 21872 utilized classical culture-based isolation techniques coupled with biochemical confirmation steps. The work also considered simplification of the biochemical confirmation steps. In addition, because of advances in molecular based methods for identification of human pathogenic Vibrio spp. classical and real-time PCR options were also included within the scope of the validation. These considerations formed the basis of a multi-laboratory validation study with the aim of improving the precision of this ISO technical specification and providing a single ISO standard method to enable detection of these important foodborne Vibrio spp.. To achieve this aim, an international validation study involving 13 laboratories from 9 countries in Europe was conducted in 2013. The results of this validation have enabled integration of the two existing technical specifications targeting the detection of the major foodborne Vibrio spp., simplification of the suite of recommended biochemical identification tests and the introduction of molecular procedures that provide both species level identification and discrimination of putatively pathogenic strains of V. parahaemolyticus by the determination of the presence of theromostable direct and direct related haemolysins. The method performance characteristics generated in this have been included in revised international standard, ISO 21872:2017, published in July 2017.

A Real-Time Recombinase Polymerase Amplification Method for Rapid Detection of Vibrio vulnificus in Seafood

“…The conventional methods for the detection of V. vulnificus usually include pre-enrichment, isolation, and biochemical identifications, which are time-consuming and labor-intensive ( Hartnell et al, 2019 ). Moreover, the culture-based methods are not sensitive to the viable but non-culturable (VBNC) status of the bacterium, and other bacteria with similar biochemical characteristics interfere ( O’Hara et al, 2003 ; Li et al, 2014 ).…”

Detection of Vibrio vulnificus in Seafood With a DNAzyme-Based Biosensor

“…At present, the detection of V. vulnificus is mainly performed by traditional culture methods (Hartnell et al, 2018), such as plate counting and the MPN method, both of which entail complex processes such as overnight culture, selective plate separation, biochemical identification, and serological experiments. These procedures are not only time consuming and labor intensive but are also prone to contamination with other bacteria in the sample, which may interfere with the identification of target bacteria.…”

Rapid and Sensitive Detection of Vibrio vulnificus Using CRISPR/Cas12a Combined With a Recombinase-Aided Amplification Assay

“…The traditional methods for detection of V. vulnificus are laborious, time-consuming, and even false positive ( O’Hara et al, 2003 ; Hartnell et al, 2019 ), which is obviously not suitable for early diagnosis and on-site detection; thus, they are gradually being replaced by simpler and faster nucleic acid amplification technology (NAT) comprising thermocycler-dependent NAT and thermocycler-independent (isothermal) NAT ( Asiello and Baeumner, 2011 ; El Sheikha et al, 2018 ). In thermocycler-dependent NAT, quantitative PCR (qPCR) assay has been widely used in V. vulnificus detection ( Campbell and Wright, 2003 ; Panicker and Bej, 2005 ).…”