Specific Discrimination of Three Pathogenic Salmonella enterica subsp. enterica Serotypes by carB -Based Oligonucleotide Microarray

Abstract

It is important to rapidly and selectively detect and analyze pathogenic Salmonella enterica subsp. enterica in contaminated food to reduce the morbidity and mortality of Salmonella infection and to guarantee food safety. In the present work, we developed an oligonucleotide microarray containing duplicate specific capture probes based on the carB gene, which encodes the carbamoyl phosphate synthetase large subunit, as a competent biomarker evaluated by genetic analysis to selectively and efficiently detect and discriminate three S. enterica subsp. enterica serotypes: Choleraesuis, Enteritidis, and Typhimurium. Using the developed microarray system, three serotype targets were successfully analyzed in a range as low as 1.6 to 3.1 nM and were specifically discriminated from each other without nonspecific signals. In addition, the constructed microarray did not have cross-reactivity with other common pathogenic bacteria and even enabled the clear discrimination of the target Salmonella serotype from a bacterial mixture. Therefore, these results demonstrated that our novel carB-based oligonucleotide microarray can be used as an effective and specific detection system for S. enterica subsp. enterica serotypes.

Discrimination of bacteriophage infected cells using locked nucleic acid fluorescent in situ hybridization (LNA-FISH)

“…On average, for a phage belonging to the Podoviridae family, 80-300 progeny phages are released upon lysis of an infected bacterium (Shin et al 2014). Also, to assemble each particle, more than 1,000 copies of major capsid proteins need to be produced (Miller et al 2003).…”

Comparing the ability of luminex xMAP®salmonella serotyping assay and traditional serotyping method for serotyping salmonella isolated from southern Chinese population

“…; Shin et al . ). The microarray chip is prohibitively expensive and less cost‐effective since it is restricted to one sample per slide.…”

“…Moreover, it was difficult to provide backward-compatible historical epidemiologic data. Microarray technology is being developed to characterize Salmonella serotypes (Guo et al 2013;Shin et al 2014). The microarray chip is prohibitively expensive and less cost-effective since it is restricted to one sample per slide.…”

Multiplex 16S rRNA‐derived geno‐biochip for detection of 16 bacterial pathogens from contaminated foods

“…Indeed, it might need to employ novel biomarkers such as virulence factor genes for more specific detections of Salmonella , Shigella , and E. coli . Previously, we conducted the study for specific discrimination of three Salmonella serotypes with a new biomarker gene . However, because the use of such biomarker gene is restricted to detection of a particular bacterium, the 16S rRNA‐derived geno‐biochip might be more suitable for multiple detection of various bacterial species in food samples.…”