PCR and oligonucleotide array for detection of Enterobacter sakazakii in infant formula

Liu, Yin; Gao, Qili; Zhang, Xia; Hou, Yanmei; Yang, Jinliang; Huang, Xi‐Tai

doi:10.1016/j.mcp.2005.08.004

Cited by 65 publications

(39 citation statements)

References 23 publications

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“…Recently, a number of molecular-based techniques for the rapid detection of Cronobacter spp. have been reported, including PCR assays (12)(13)(14), the DuPont Qualicon BAX system (15), real-time PCR (5,8,11), and oligonucleotide arrays (14). Real-time PCR assays using the TaqMan probe have been increasingly exploited, and these assays have replaced ordinary PCR methods in many cases.…”

Section: Resultsmentioning

confidence: 99%

Real-time PCR targeting OmpA gene for detection of Cronobacter spp. in powdered infant formula

Dong¹,

Wu²,

Wu³

et al. 2013

Food Sci Biotechnol

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Cronobacter spp. (formerly Enterobacter sakazakii), a pathogen commonly found in powdered infant formula (PIF), is a rare cause of invasive infection with a high mortality rate in neonates. In the present study, a realtime PCR assay was conducted to identify the pathogens in PIF using a TaqMan probe targeting the outer membrane protein A gene (ompA) of Cronobacter spp. The specificity of the PCR assay was tested against 25 strains of Cronobacter spp. and 38 non-Cronobacter bacterial species. The detection limits of this method are 1.0×10 2 copy/µL in standard plasmid, 1.1 CFU/100 g in PIF through 38 h of enrichment, and 2.8×102 CFU/mL in phosphate-buffered saline (pH 7.0). Based on the detection limits, real-time PCR is more sensitive than simplex and duplex PCR. These methods were successfully applied to actual samples, indicating that this real-time PCR assay can be used for the detection of Cronobacter spp. in PIF.

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Section: Resultsmentioning

confidence: 99%

Real-time PCR targeting OmpA gene for detection of Cronobacter spp. in powdered infant formula

Dong¹,

Wu²,

Wu³

et al. 2013

Food Sci Biotechnol

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show abstract

“…Blue-green colonies were picked and subjected to further biochemical and molecular confirmative techniques. Then the 16S rRNA gene of the isolates was amplified using two bacterial universal primers, 27F and 1492R, and further confirmed with species-specific PCR to detect the 16S-23S rDNA internal transcribed spacer (ITS) (Liu et al 2006). The similarity test of the 16S rRNA gene sequence was performed by using BLAST program on NCBI http://www.ncbi.nlm.nih.gov.…”

Section: Methodsmentioning

confidence: 99%

Isolation of Cronobacter spp. isolates from infant formulas and their survival in the production process of infant formula

Fu¹,

Gao²,

Liu³

et al. 2011

Czech J. Food Sci.

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Fu S. , Gao J., Liu Y., Chen H. (2011): Isolation of Cronobacter spp. isolates from infant formulas and their survival in the production process of infant formula. Czech J. Food Sci., 29: 391-399.Over a 24-month surveillance, three Cronobacter strains, NC041, NC830, and NC1006, were isolated from 77 powder infant formulas (3.90%). No Cronobacter was detected in liquid milk. The prevalence of Cronobacter in the prefinal product and packaged final product was 3.70% and 4.35%, respectively. The isolated Cronobacter strains were subjected to several lethal challenges including the pH, drying, disinfectant, and simulated infant formulas manufacturing process (SIFMP). The results indicated that they exhibited unusual resistance to the dry stress and disinfectant. In SIFMP, Cronobacter isolates were inoculated into three possible contamination entry points involving the stages prior to heating, drying, and filling, respectively. No Cronobacter could survive the heating. However, a high level (10 5 CFU/ml) of Cronobacter was detected in the samples after the inoculation at the drying point. Furthermore, the survival of Cronobacter was observed during the storage at 10°C after 7 days. The results support the hypothesis that Cronobacter contamination occurs at the stages of drying and filling, and highlight the need to improve the disinfecting measures in the manufacturing process.

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“…sakazakii in 81 dry food samples was described by Liu et al (2006). The preparation of templates and PCR procedure was previously described.…”

Section: Enrichment Procedures Of Commercial Food Samples and Checkingmentioning

confidence: 99%

Analysis of major band of Enterobacter sakazakii by ERIC-PCR and development of a species-specific PCR for detection of Ent. sakazakii in dry food samples

Zhou

et al. 2008

Journal of Microbiological Methods

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PCR and oligonucleotide array for detection of Enterobacter sakazakii in infant formula

Cited by 65 publications

References 23 publications

Real-time PCR targeting OmpA gene for detection of Cronobacter spp. in powdered infant formula

Real-time PCR targeting OmpA gene for detection of Cronobacter spp. in powdered infant formula

Isolation of Cronobacter spp. isolates from infant formulas and their survival in the production process of infant formula

Analysis of major band of Enterobacter sakazakii by ERIC-PCR and development of a species-specific PCR for detection of Ent. sakazakii in dry food samples

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