%0 Journal Article
%T A robotic DNA purification protocol and real-time PCR for the detection of Enterobacter sakazakii in powdered infant formulae
%A Sylviane Derzelle
%A Fran£żoise Dilasser
%J BMC Microbiology
%D 2006
%I BioMed Central
%R 10.1186/1471-2180-6-100
%X The real-time PCR assay had 100% specificity as assessed using 35 E. sakazakii and 184 non-E. sakazakii strains. According to the E. sakazakii strains tested, the detection limits ranged from 5 to 25 genomic copies. Assays on pure cultures (including real-time PCR and DNA extraction) gave a sensitivity of about 102 to 103 CFU/ml. Out of 41 naturally contaminated infant formulae and environmental samples analysed for the presence of E. sakazakii, 23 were positive by real-time PCR and 22 by the conventional culture method, giving 97.5% concordance with the ISO-IDF reference method.This method, combining specific real-time PCR, automated DNA extraction and ISO-IDF standard enrichments, provides a useful tool for rapid screening of E. sakazakii in food and environmental matrices.Enterobacter sakazakii, previously known as yellow-pigmented E. cloacae [1], has been identified as the causative agent of rare but often severe invasive infections in infants. Neonates and infants less than 2 months of age appear to be groups at particular risk [2,3]. Meningitis is the most frequently reported clinical symptom in neonatal E. sakazakii infections [2,4-7]. Infants in whom meningitis developed were generally < 1 week of age at the onset of infection and had near-term gestational age and birthweight [3]. They frequently develop complications including seizures and brain abscesses, resulting in high mortality (over 40%) and morbidity. Septicaemia or necrotizing enterocolitis are also associated with E. sakazakii-related infections [2,4-7]. Compared with infants with meningitis, infants in whom bacteremia alone developed tended to be born very prematurely and have extremely low birthweight (<1,000 g). They had generally surpassed the neonatal period at the onset of their disease [3]. Mortality among these cases is lower (about 10%) [2,4-7].E. sakazakii is emerging as a hazard in powdered infant formulae (PIF). In several E. sakazakii-related outbreaks and sporadic cases, PIF was epid
%U http://www.biomedcentral.com/1471-2180/6/100