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 En...

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Bibliographic Details
Published in:International journal of food microbiology 2007-05, Vol.116 (2), p.214-220
Main Authors: Hassan, Abdulwahed Ahmed, Akineden, Ömer, Kress, Claudia, Estuningsih, Sri, Schneider, Elisabeth, Usleber, Ewald
Format: Article
Language:English
Subjects:
16S rRNA
bacterial contamination
Base Sequence
Biological and medical sciences
Citrobacter koseri
Consumer Product Safety
Cronobacter sakazakii
Cronobacter sakazakii - genetics
Cronobacter sakazakii - isolation & purification
DNA, Bacterial - chemistry
Enterobacter
Enterobacter sakazakii
Enterobacteriaceae
food contamination
Food Contamination - analysis
Food industries
Food microbiology
food pathogens
Fundamental and applied biological sciences. Psychology
genes
Genetic Variation
Humans
Infant
Infant Formula
infant formulas
Infant, Newborn
molecular sequence data
nucleotide sequences
pathogen identification
PCR
polymerase chain reaction
Polymerase Chain Reaction - methods
powdered foods
Powdered infant formula
ribosomal RNA
ribotypes
RNA, Ribosomal, 16S - genetics
Salmonella enterica
Sequence Homology, Nucleic Acid
Species Specificity
virulence
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Summary: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.
ISSN:0168-1605
1879-3460
DOI:10.1016/j.ijfoodmicro.2006.12.011