Cronobacter Support
05-19-2009, 02:46 PM
1st International Conference on Cronobacter Poster Abstract 41
Express design and manufacturing of E. sakazakii TaqMan Pathogen Detection Kit by Applied Biosystems
E. sakazakii (now re-named Cronobacter) is a gram-negative rod-shaped bacterium which is a rare cause of invasive infection with high mortality rates in neonates. Powdered milk-based infant formulas have been associated with the E. sakazakii-related outbreaks in premature or other immunocompromised infants. In general, the reported case-fatality rate varies from 40-80 % among newborns diagnosed with this type of severe infection. The type of meningitis caused by E. sakazakii may lead to cerebral abscess or infarction with cyst formation and severe neurologic impairment. The conventional method approved by FDA requires 5 days for detection. To improve sensitivity and shorten detection time Applied Markets Division has designed a TaqMan Pathogen Detection Kit for E. sakazakii. Working on alternatives to 16S-based TaqMan design, we selected RnaseP gene as a good TaqMan target based on literature and our previous experience. However, there is no RnaseP sequence for E. sakazakii in the public databases. Based on Bioinformatics analysis, we selected conserved regions across Enterobacter species to design primers to amplify, and sequence RnaseP gene in ATCC E. sakazakii strains. Using the new sequence information we designed a real-time PCR assay to specifically detect E. sakazakii. The assay was tested with four target strains and a panel of antitargets including Salmonella enterica, Shigella flexneri, Escherichia coli, Enterobacter nimipressuralis, Enterobacter dissolvens, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundi and Listeria monocytogenes.
Cristina Rodriquez
Ordesa, Spain.
Express design and manufacturing of E. sakazakii TaqMan Pathogen Detection Kit by Applied Biosystems
E. sakazakii (now re-named Cronobacter) is a gram-negative rod-shaped bacterium which is a rare cause of invasive infection with high mortality rates in neonates. Powdered milk-based infant formulas have been associated with the E. sakazakii-related outbreaks in premature or other immunocompromised infants. In general, the reported case-fatality rate varies from 40-80 % among newborns diagnosed with this type of severe infection. The type of meningitis caused by E. sakazakii may lead to cerebral abscess or infarction with cyst formation and severe neurologic impairment. The conventional method approved by FDA requires 5 days for detection. To improve sensitivity and shorten detection time Applied Markets Division has designed a TaqMan Pathogen Detection Kit for E. sakazakii. Working on alternatives to 16S-based TaqMan design, we selected RnaseP gene as a good TaqMan target based on literature and our previous experience. However, there is no RnaseP sequence for E. sakazakii in the public databases. Based on Bioinformatics analysis, we selected conserved regions across Enterobacter species to design primers to amplify, and sequence RnaseP gene in ATCC E. sakazakii strains. Using the new sequence information we designed a real-time PCR assay to specifically detect E. sakazakii. The assay was tested with four target strains and a panel of antitargets including Salmonella enterica, Shigella flexneri, Escherichia coli, Enterobacter nimipressuralis, Enterobacter dissolvens, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundi and Listeria monocytogenes.
Cristina Rodriquez
Ordesa, Spain.