[Development and validation of a multiplex polymerase chain reaction for molecular identification of Salmonella enterica serogroups B, C2, D and E].
BACKGROUND
The scheme Kauffman-White (KW) for serotyping of Salmonella recognizes 46 O antigens, and 119 H antigens, thereby permitting the characterization of 2541 serotypes. The serotyping is a useful epidemiological tool in identifying circulating serotypes and to characterize outbreaks. However, the method presents technical limitations, difficulty in interpretation of results and high costs.


OBJECTIVE
A multiplex polymerase chain reaction test (M-PCR) was developed as an alternative method for the identification of serogroups B, C2, D, and E of Salmonella enterica.


METHODS
The M-PCR detected Salmonella genes rfbJ of serogroups B and C2 and wzx of serogroups D and E. To standardize the M-PCR, reference strains of Salmonella serogroups were compared. Amplification of invA gender-specific gene of Salmonella was included as internal control of amplification. To validate the test, a blind study was conducted to identify by M-PCR 400 isolates that had been previously characterized by serology.


RESULTS
The M-PCR detected Salmonella serogroups with reproducible results (Kappa index = 0.95). The sensitivity of the test was between 98% to 100% and specificity between 96% to 100%.


CONCLUSIONS
The polymorphisms in the Salmonella genes rfbJ and wzx permitted the development of a method for molecular typing of Salmonella serogroups that was sensitive, specific and reproducible.
Universidad Nacional de Colombia, Medellín, Colombia.
Lelia
L
Lavalett
Miryan Margot
MM
Sánchez
Nélida
N
Múñoz
Jaime
J
Moreno
Nora
N
Cardona-Castro
D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D006801:Humans; D019081:O Antigens / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods* / Q000592:standards; D015203:Reproducibility of Results; D019779:Salmonella enterica / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000191:economics; D016037:Single-Blind Method
D000914:Antibodies, Viral; D000956:Antigens, Viral; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D019081:O Antigens; C075518:invA protein, Bacteria