Development and validation of a PulseNet standardized protocol for subtyping isolates of Cronobacter species.
Cronobacter (formerly known as Enterobacter sakazakii) is a genus comprising seven species regarded as opportunistic pathogens that can be found in a wide variety of environments and foods, including powdered infant formula (PIF). Cronobacter sakazakii, the major species of this genus, has been epidemiologically linked to cases of bacteremia, meningitis in neonates, and necrotizing enterocolitis, and contaminated PIF has been identified as an important source of infection. Robust and reproducible subtyping methods are required to aid in the detection and investigation, of foodborne outbreaks. In this study, a pulsed-field gel electrophoresis (PFGE) protocol was developed and validated for subtyping Cronobacter species. It was derived from an existing modified PulseNet protocol, wherein XbaI and SpeI were the primary and secondary restriction enzymes used, generating an average of 14.7 and 20.3 bands, respectively. The PFGE method developed was both reproducible and discriminatory for subtyping Cronobacter species.
Servicio Enterobacterias, Departamento Bacteriología, INEI-ANLIS Dr. Carlos G. Malbrán, Buenos Aires, Argentina.
Silvina P
SP
Brengi
Stephen B
SB
O'Brien
Mariana
M
Pichel
Carol
C
Iversen
Matthew
M
Arduino
Norma
N
Binsztein
Bette
B
Jensen
Franco
F
Pagotto
Efrain M
EM
Ribot
Roger
R
Stephan
Nicole
N
Cernela
Kara
K
Cooper
Séamus
S
Fanning
D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D059126:Cronobacter / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D044083:Cronobacter sakazakii / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D004756:Enterobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology; D057141:Food, Preserved / Q000382:microbiology; D005517:Foodborne Diseases / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D008892:Milk / Q000382:microbiology; D058889:Molecular Typing / Q000379:methods*; D015203:Reproducibility of Results; D014734:Vibrio cholerae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015010:Yersinia pestis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes