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Journal of Clinical Microbiology, October 2009, p. 3178-3184, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00366-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Detection of Enterobacter sakazakii and Other Pathogens Associated with Infant Formula Powder by Use of a DNA Microarray{triangledown} ,{dagger}

Min Wang,1,2,3,4,{ddagger} Boyang Cao,1,2,3,4,{ddagger} Qili Gao,6 Yamin Sun,1,2,3,4 Pei Liu,6 Lu Feng,1,2,3,4,5 and Lei Wang1,2,3,4,5*

TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 HongDa Street, TEDA, Tianjin 300457, China,1 Key Laboratory of Molecular Microbiology and Technology of the Ministry of Education, Nankai University, Tianjin 300071, China,2 Tianjin State Laboratory of Microbial Functional Genomics, 23 HongDa Street, TEDA, Tianjin 300457, China,3 Tianjin Research Center for Functional Genomics and Biochips, 23 HongDa Street, TEDA, Tianjin 300457, China,4 Tianjin Biochip Corporation, 23 HongDa Street, TEDA, Tianjin 300457, China,5 Tianjin Entry-Exit Inspection and Quarantine Bureau, 8 ZhaoFa Residential Quarter, 2nd St., TEDA, Tianjin 300457, China6

Received 18 February 2009/ Returned for modification 4 May 2009/ Accepted 23 July 2009

Pathogen detection is critical to the process of generating and testing powdered infant formula (PIF). An obstacle associated with PIF microbial surveillance is that most current procedures are time-consuming and labor-intensive. We have developed a rapid, DNA microarray-based detection technique to identify 10 different pathogenic bacteria associated with PIF contamination based on the 16S-23S rRNA gene internal transcribed spacer (ITS) sequences and wzy (O antigen polymerase) gene. Using this procedure, Enterobacter sakazakii, Salmonella enterica, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Acinetobacter baumannii, Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli O157 were identified. One hundred eighty-five strains were used to validate the microarray assay (including 134 target pathogen strains and 51 closely related bacteria). Twenty-seven probes reproducibly detected multiple pathogens with high specificity and sensitivity (0.100 ng genomic DNA or 104 CFU/ml). Twenty-one real PIF samples were tested by the microarray with 100% accuracy. The data presented reveal that the designed oligonucleotide microarray is a promising method for basic microbiology, clinical diagnosis, food safety, and epidemiological surveillance.

* Corresponding author. Mailing address: TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 HongDa St., TEDA, Tianjin 300457, China. Phone: 86-22-66229588. Fax: 86-22-66229596. E-mail: wanglei{at}nankai.edu.cn

{triangledown} Published ahead of print on 29 July 2009.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

{ddagger} These authors contributed equally to this report.

Journal of Clinical Microbiology, October 2009, p. 3178-3184, Vol. 47, No. 10
0095-1137/09/$08.00+0     doi:10.1128/JCM.00366-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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