A new multiplex PCR for differential identification of Shigella flexneri and Shigella sonnei and detection of Shigella virulence determinants.
Most of the multiplex PCR (mPCR) used to identify Shigella do not discriminate between Shigella species or serotypes. We designed a mPCR to differentiate between S. flexneri and S. sonnei strains based on the detection of markers associated with the she pathogenicity island described in Shigella. In addition, specific primers were included to detect the Shigella virulence determinants ShET-1 and ShET-2 enterotoxin genes. The analysis of 304 Shigella strains from Chile and 79 Shigella strains from other geographic locations indicated that the mPCR described here detected all Shigella species and specifically differentiated S. flexneri and S. sonnei. The technique was sensitive, reproducible, specific and simple to perform, providing a new tool with the potential to be employed for epidemiological and diagnostic purposes.
Farfán
M J
MJ
Departamento de Pediatría, Hospital Dr Luis Calvo Mackenna, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
Garay
T A
TA
Prado
C A
CA
Filliol
I
I
Ulloa
M T
MT
Toro
C S
CS
D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D002648:Child; D002675:Child, Preschool; D002677:Chile; D004269:DNA, Bacterial / Q000235:genetics; D004405:Dysentery, Bacillary / Q000175:diagnosis / Q000382:microbiology*; D004768:Enterotoxins / Q000235:genetics; D044404:Genomic Islands; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012763:Shigella flexneri / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012764:Shigella sonnei / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D037521:Virulence Factors / Q000235:genetics*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004768:Enterotoxins; D037521:Virulence Factors