A TaqMan Based Real-Time PCR Assay with the Internal Amplified Control for the Detection of Enterobacter sakazakii in Infant Formula

2009 ◽  
Author(s):  
Xudong Su ◽  
Chao Zhang ◽  
Xiaoyan Ma ◽  
Wei Zhang ◽  
Yingjun Li ◽  
...  
Keyword(s):  
Real Time ◽  
Infant Formula ◽  
Real Time Pcr ◽  
Enterobacter Sakazakii ◽  
Pcr Assay

Rapid, Specific Detection of Enterobacter sakazakii in Infant Formula Using a Real-Time PCR Assay

2005 ◽  
Vol 68 (1) ◽  
pp. 59-63 ◽  
Author(s):  
K. H. SEO ◽  
R. E. BRACKETT
Keyword(s):  
Real Time ◽  
Infant Formula ◽  
Real Time Pcr ◽  
Regulatory Agencies ◽  
Invasive Infection ◽  
Enterobacter Sakazakii ◽  
Specific Detection ◽  
Pcr Assay ◽  
Powdered Milk ◽  
Food Industries

Enterobacter sakazakii 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 this study, an assay was developed for the specific detection of E. sakazakii in infant formula using an application of the fluorogenic 5′ nuclease assay (TaqMan). A set of primers and probe was designed using the E. sakazakii partial macromolecular synthesis operon: the rpsU gene 3′ end and the primase (dnaG) gene 5′ end. The specificity of the assay was evaluated using 68 Enterobacter and 55 non-Enterobacter strains. The newly developed assay enables us to detect 100 CFU/ml in pure culture and in reconstituted infant formula in 50 cycles of PCR without enrichment. The assay was specific enough to discriminate E. sakazakii from all other Enterobacter and non-Enterobacter strains tested. The developed real-time PCR assay could save up to 5 days and eliminate the need for plating samples on selective or diagnostic agars and for biochemical confirmation steps. The real-time PCR assay could be used to rapidly screen infant formula samples for E. sakazakii and would be a boon to food industries and regulatory agencies.

Real time PCR using TaqMan and SYBR Green for detection of Enterobacter sakazakii in infant formula

2006 ◽  
Vol 65 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Yin Liu ◽  
Xiaoning Cai ◽  
Xia Zhang ◽  
Qili Gao ◽  
Xiaochuan Yang ◽  
...  
Keyword(s):  
Real Time ◽  
Infant Formula ◽  
Real Time Pcr ◽  
Sybr Green ◽  
Enterobacter Sakazakii

Development of an Improved Protocol for the Isolation and Detection of Enterobacter sakazakii (Cronobacter) from Powdered Infant Formula

2010 ◽  
Vol 73 (6) ◽  
pp. 1016-1022 ◽  
Author(s):  
YI CHEN ◽  
KWANG-YONG SONG ◽  
ERIC W. BROWN ◽  
KEITH A. LAMPEL
Keyword(s):  
Real Time ◽  
Infant Formula ◽  
Real Time Pcr ◽  
Powdered Infant Formula ◽  
Enterobacter Sakazakii ◽  
Biochemical Tests ◽  
The Real

Enterobacter sakazakii causes severe maladies and, in some cases, is fatal among infants. Powdered infant formula (PIF) contaminated with E. sakazakii has been documented as a potential cause of several outbreaks involving infants. This study describes the development of a method for the isolation and detection of E. sakazakii from PIF. It combines Taqman real-time PCR, Brilliance E. sakazakii and R&F chromogenic agars, and RAPID ID 32E biochemical tests. This method provides an expedient analysis within 1 to 2 days depending on the amount and stress status of E. sakazakii organisms and competing microorganisms in PIF. The real-time PCR has bifunctional applications, including both screening and culture confirmation of E. sakazakii.

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