Identification by a multiplex PCR-based assay of Salmonella typhimurium and Salmonella enteritidis strains from environmental swabs of poultry houses.
A multiplex-PCR-based assay (m-PCR) was developed for the detection of Salmonella and for the identification of the two serotypes Enteritidis and Typhimurium. Three sets of primers selected from different genomic sequences amplified a 429 bp fragment specific for the genus Salmonella within a randomly cloned sequence, a 559 bp target specific for Salmonella Typhimurium within the fliC gene and a 312 bp fragment specific for Salmonella Enteritidis within the sefA gene. The m-PCR-based assay was used for detecting Salmonella from 1078 environmental swabs of poultry houses. Prior to PCR, these swabs were pre-enriched in phosphate-buffered peptone water for 18-20 h and then sub-cultured on a Modified Semi-solid Rappaport Vassiliadis medium (MSRV) for 18-20 h. The m-PCR combined with MSRV had a better sensitivity (95%) than the bacteriological method (92.5%). The MSRV-m-PCR assay and the bacteriological method had an agreement rate of 95.6%.
Soumet
C
C
Unité de Recherche en Hygiène et Qualité des Produits Avicoles et Porcins. c.soumet@fougeres.cneva.fr
Ermel
G
G
Rose
V
V
Rose
N
N
Drouin
P
P
Salvat
G
G
Colin
P
P
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques; D002645:Chickens; D017931:DNA Primers; D004783:Environmental Microbiology; D036781:Fimbriae Proteins; D005408:Flagellin / Q000235:genetics; D006799:Housing, Animal; D016133:Polymerase Chain Reaction / Q000379:methods*; D012477:Salmonella enteritidis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D001426:Bacterial Proteins; D017931:DNA Primers; C080931:sefA protein, Salmonella enteritidis; D005408:Flagellin; D036781:Fimbriae Proteins