Characterization of the gene encoding the 16S rRNA of Enterobacter sakazakii and development of a species-specific PCR method.
The gene encoding the 16S rRNA of Enterobacter (E.) sakazakii (ATCC 29544, plus four strains isolated from powdered infant formula) was studied, and the sequence compared with those of other Enterobacteriaceae in aspects of genetic variability. Sequence differences between E. sakazakii and other Enterobacteriaceae within the hypervariable regions V1, V2, and V3, respectively, were used to develop two PCR methods for E. sakazakii. PCR1 employed a primer pair located in V1/V2, while PCR2 utilized a primer pair located in V1/V3, respectively. The two PCR methods were tested against a set of 57 E. sakazakii and 148 non-E. sakazakii isolates. PCR1 gave an amplicon with a size of 406 bp and resulted in 100% positive results for E. sakazakii, but also detected Citrobacter koseri/amalonaticus and all nine tested Salmonella enterica serovars. In contrast, PCR2 (amplicon size of 952 bp) gave positive results only for E. sakazakii, thus allowing specific identification of this species.
Hassan
Abdulwahed Ahmed
AA
Dairy Sciences, Institute of Veterinary Food Science, Justus-Liebig-University, Ludwigstrasse 21, Giessen, Germany. abdulwahed.hassan@vetmed.uni-giessen.de
Akineden
Omer
O
Kress
Claudia
C
Estuningsih
Sri
S
Schneider
Elisabeth
E
Usleber
Ewald
E
D001483:Base Sequence; D003257:Consumer Product Safety; D044083:Cronobacter sakazakii / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000737:chemistry; D005506:Food Contamination / Q000032:analysis*; D014644:Genetic Variation; D006801:Humans; D007223:Infant; D041943:Infant Formula; D007231:Infant, Newborn; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S