Rapid identification of Salmonella enterica serovars, Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum, by multiplex PCR.

Salmonella enterica subsp. enterica poses a threat to both human and animal health, with more than 2500 serovars having been reported to date. Salmonella serovars are identified by slide and tube agglutination tests using O and H antigen-specific anti-sera, although this procedure is both labor intensive and time consuming. Establishment of a method for rapid screening of the major Salmonella serovars is therefore required. We have established multiplex polymerase chain reaction (m-PCR) assays for identification of seven serovars of Salmonella, i.e., Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum. Three serovar-specific genomic regions (SSGRs) of each serovar were selected using an approach in comparative genomics. The Salmonella-specific invA gene was used to confirm the genetic background of the organisms. The isolates tested were identified as a target serovar when the three selected SSGRs and invA were all positive for amplification. The specificity of each m-PCR assay was investigated using 118 serovars of Salmonella and 12 species of non-Salmonella strains. Although a small number of false-positive results were observed in the m-PCR assays used to identify Typhimurium, Choleraesuis, Enteritidis and Dublin for closely related serovars, false-negative results were not observed in any assays. These assays had sufficient specificity to identify the seven Salmonella serovars, and therefore, have the potential for use as rapid screening methods.

Akiba M ，Kusumoto M ，Iwata T 《-》

Identification of Salmonella enterica subspecies I, Salmonella enterica serovars Typhimurium, Enteritidis and Typhi using multiplex PCR.

Park SH ，Kim HJ ，Cho WH ，Kim JH ，Oh MH ，Kim SH ，Lee BK ，Ricke SC ，Kim HY ... -  《-》

Detection and identification of Salmonella Typhimurium in bovine diarrhoeic fecal samples by immunomagnetic separation and multiplex PCR assay.

Salehi TZ ，Tadjbakhsh H ，Atashparvar N ，Nadalian MG ，Mahzounieh MR ... -  《Zoonoses and Public Health》

Validation of a seminested PCR approach for rapid detection of Salmonella enterica subsp. enterica serovar Gallinarum.

Salmonella enterica subsp. enterica serovar Gallinarum (S. Gallinarum) is the causative agent of fowl typhoid, one of the major causes of mortality and morbidity on poultry farms. Even though it has been substantially eradicated in many developed countries, the disease still remains endemic in Central and South America, in Africa and in the Mediterranean countries of Europe. This leads to the routine screening of flocks, mainly by cultivation and serological techniques, which are expensive, as well as time and labour-consuming. Here we describe a simple and specific PCR-based method for detecting S. Gallinarum. It relies on two seminested PCRs which use four pairs of primers designed on the basis of two genomic regions which appear to be exclusive to the pathogen. Furthermore, an internal positive control was devised in order to avoid any false negative results. We performed sensitivity and specificity tests, and our findings showed the cogency of the system and its potential effectiveness even for routine uses.

Pugliese N ，Circella E ，Pazzani C ，Pupillo A ，Camarda A ... -  《-》

A novel real-time polymerase chain reaction for identification of Salmonella enterica subspecies enterica.

Salmonella enterica subspecies enterica (subspecies I) causes the majority of infections in humans and homeothermic animals. We present a real-time polymerase chain reaction assay targeting the hilA gene that demonstrates 97.9% specificity and 99.9% sensitivity for rapid and reliable identification of subspecies I, offering savings in time and labor over traditional methods.

Hopkins KL ，Lawson AJ ，Connell S ，Peters TM ，de Pinna E ... -  《-》