Detection of Chromobacterium violaceum by multiplex PCR targeting the prgI, spaO, invG, and sipB genes.
Based on the recently completed genomic sequence of Chromobacterium violaceum American Type Culture Collection (ATCC) 12472 a multiplex PCR assay targeting the prgI, spaO, invG, and sipB genes of the Salmonella SPI-1 homologue type-III secretion system was developed. PCR products of 255bp (prgI), 749bp (spaO), 1685bp (invG), and 1752bp (sipB) were successfully amplified simultaneously in a single reaction with all Chr. violaceum strains investigated whereas other bacteria tested negative. The detection limit for pure cultures in multiplex PCR analysis was 100CFU. The developed assay significantly improves rapid identification of Chr. violaceum and allows its differentiation from closely related organisms.
Scholz
Holger Christian
HC
Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, D-80937 Munich, Germany. Holger1Scholz@bundeswehr.org
Witte
Angela
A
Tomaso
Herbert
H
Al Dahouk
Sascha
S
Neubauer
Heinrich
H
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D002861:Chromobacterium / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C101106:SpaO protein, Salmonella sp.; C090991:invG protein, Salmonella typhimurium; C094579:invasion protein B, Salmonella typhimurium