Abstract
Salmonellosis is responsible for large number of infections in both human and animals. Salmonella enterica serovar Typhi is a causative agent of typhoid fever and Salmonella enterica serovar Paratyphi A is a causative agent of paratyphoid fever. Conventional methods of isolation of Salmonella strains take 4-6 days to complete and are therefore laborious and require substiantial manpower. Therefore development of a PCR assay that can target multiple genes for rapid detection of S. Typhi and S.Paratyphi A. Methods: Synthetic primers for O, H, and Vi antigen genes, tyv , prt , fliC-d, fliC-a, and viaB, were used for the rapid identification of S. Typhi and Paratyphi A with Multiplex PCR. Results: All the clinical isolates examined were accurately identified by this PCR technique, that differentiated S. Typhi and Paratyphi A, based on size and number of amplified fragments. S. enterica serovar Typhi, yielded four bands of tyv(tyvelose epimerase gene, 615bp),prt (paratose gene, 258bp),flic-d(phage-1 flagellin gene for d- antigen 750bp) and viaB ( vi antigen gene, 439bp); S.enterica serovar Paratyphi A yielded only two bands prt (paratose gene, 258bp) and flic-a (phage-1 flagellin gene for a- antigen 329bp). Conclusion: These data indicate that multiplex PCR is a potentially valuable tool for rapid diagnosis of Salmonella enterica serovar Typhi and Paratyphi A from clinical isolates.
Highlights
750bp) and viaB ( vi antigen gene, 439bp); S.enterica serovar Paratyphi A yielded only two bands prt and flic-a. These data indicate that multiplex Polymerase chain reaction (PCR) is a potentially valuable tool for rapid diagnosis of Salmonella enterica serovar Typhi and Paratyphi A from clinical isolates
Materials and methods Bacterial strains: The study included six isolates of Salmonella enteric serovar Typhi and three isolates of Salmonella enterica serovar Paratyphi A obtained in 2007 from the Central Public Health Laboratory (CPHL)- Enteric Department in Baghdad and diagnosed as Salmonella using conventional methods by biochemical tests, Kuffman and white scheme was followed for serological confirmation and typing using Anti-Salmonella sera (Biomerieux)
Different isolates were used in this study provided from CPHL for the specificity test required for Multiplex-PCR, included Salmonella Enteritidis, Salmonella Typhimurium, and Proteus mirablis
Summary
750bp) and viaB ( vi antigen gene, 439bp); S.enterica serovar Paratyphi A yielded only two bands prt (paratose gene, 258bp) and flic-a (phage-1 flagellin gene for a- antigen 329bp). Conclusion: These data indicate that multiplex PCR is a potentially valuable tool for rapid diagnosis of Salmonella enterica serovar Typhi and Paratyphi A from clinical isolates. Others have already used( Vi antigen gene , H antigen genes , and O antigen synthesis genes ), for rapid detection of typhoid fever and paratyphoid fever based on a multiplex PCR technique that detected the Vi antigen gene (viaB), H antigen genes (fliC-d and fliC-a), and O antigen synthesis genes (tyv and prt).
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