Development of PCR primers for the detection of Salmonella enterica serovar Choleraesuis based on the fliC gene.
Salmonella enterica serovar Choleraesuis may cause swine salmonellosis and human infection. Because the conventional method for detection of this Salmonella serovar may take 3 to 5 days, a PCR method for detection was evaluated. By comparing the sequence of the phase 1 flagellin (fliC) gene of Salmonella Choleraesuis with that of other Salmonella serovars and of other bacteria species available in GenBank, two PCR primers (flinC-F and flinC-R) were designed. Using these primers, all 97 Salmonella Choleraesuis strains assayed generated the expected PCR product, with a molecular mass of 963 bp. Except for S. enterica Paratyphi C, Salmonella isolates other than Salmonella Choleraesuis and non-Salmonella isolates, including strains of Enterobacteriaceae, all generated negative PCR results. Salmonella Paratyphi C could be differentiated from Salmonella Choleraesuis through the use of primers designed from the viaB gene. When Salmonella Choleraesuis isolates from swine stool, pork, liver, feed, and human whole blood samples were assayed with a preenrichment step, as low as 1 CFU/g or ml of the original sample could be detected.
Department of Food Science, National Chung-Hsing University, Taichung, Taiwan, Republic of China.
Tsai-Hsin
TH
Chiu
Jen-Chieh
JC
Pang
Wen-Zhe
WZ
Hwang
Hau-Yang
HY
Tsen
D000818:Animals; D003257:Consumer Product Safety; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D005408:Flagellin / Q000235:genetics*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D005784:Gene Amplification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D019779:Salmonella enterica / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D005408:Flagellin