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学科主题: Biochemical Research Methods; Microbiology
题名: 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
作者: Ye, YW ; Wu, QP ; Zhou, YH ; Dong, XH ; Zhang, JM
通讯作者: yeyw04@mails.gucas.ac.cn ; wuqp203@yahoo.com.cn
关键词: Enterobacter sakazakii ; ERIC (Enterobacterial Repetitive Intergenic ; Consensus)-PCR ; Infant formulas ; Major band
刊名: JOURNAL OF MICROBIOLOGICAL METHODS
发表日期: 2008
卷: 75, 期:3, 页:392-397
收录类别: sci
部门归属: [Ye, Yingwang; Wu, Qingping; Zhou, Yanhong; Dong, Xiaohui; Zhang, Jumei] Guangdong Inst Microbiol, Guangdong Prov Key Lab Microbiol Culture Collect, Guangzhou 510070, Guangdong, Peoples R China; [Ye, Yingwang; Dong, Xiaohui] Chinese Acad Sci, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Ye, Yingwang; Zhou, Yanhong; Dong, Xiaohui] Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China; [Zhou, Yanhong] Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China
项目归属: 海洋生物
摘要: ERIC (Enterobacterial Repetitive Intergenic Consensus)-PCR was employed to generate stable and reproductive ERIC-PCR fingerprints of Ent. sakazakii ATCC51329. Moreover, this study also cloned and sequenced a major band of Ent. sakazakii (ATCC51329) ERIC-PCR fingerprints. The major band was amplified with primer ERIC2 and sequences extending primer ERIC 2 showed poor similarity with ERIC elements. A comparison of the nucleotide acid with other sequences available in the GenBank revealed 90% of identity with Ent. sakazakii ATCC BAA-894, and 73%-74% of identity with oligopeptiase gene or protease gene of some species from the Enterobacteriaceae family. Two primers were synthesized to develop and optimize an Enterobacter sakazakii-specific PCR based on regions of major band unique to Ent. sakazakii. The expected fragment was amplified from all of Ent. sakazkaii but not from the negative controls. As few as 10(2) CFU/ml of Ent. sakazakii of PCR were directly detected in the infant formulas. This was the case even in the presence of other bacteria. A comparison of traditional methods and new developed PCR in commercial foods suggested that without using API20-E test, the DFI chromogenic medium and FDA method showed 46.15% and 50% false positive respectively. Moreover, one false negative was observed with FDA method. In contrast, PCR was highly sensitive and specific to Ent. sakazakii. A high heterogeneity between Ent. sakazakii and the other microorganisms was found on expected fragment sequence. In addition, Ent sakazakii ATCC51329 formed a separate branch with >5% divergence from the type strain ATCC BAA-894 and major strains. (C) 2008 Elsevier B.V. All rights reserved.
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WOS记录号: WOS:000261527900002
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内容类型: 期刊论文
URI标识: http://ir.scsio.ac.cn/handle/344004/5346
Appears in Collections:海洋生物_期刊论文

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Ye, YW; Wu, QP; Zhou, YH; Dong, XH; Zhang, JM.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,JOURNAL OF MICROBIOLOGICAL METHODS,2008,75(3):392-397
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