A PCR METHOD FOR THE DETECTION OF LISTERIA MONOCYTOGENES BASED ON A NOVEL TARGET SEQUENCE IDENTIFIED BY COMPARATIVE GENOMIC ANALYSIS
A PCR METHOD FOR THE DETECTION OF LISTERIA MONOCYTOGENES BASED ON A NOVEL TARGET SEQUENCE IDENTIFIED BY COMPARATIVE GENOMIC ANALYSIS
复制标题
基于比较基因组分析确定的新靶序列的单核细胞增生李斯特氏菌PCR检测方法
DOI：
10.1111/j.1745-4565.2010.00245.x
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发表时间：
2010-11
影响因子：
2.4
通讯作者：
Shi, Xianming
中科院分区：
文献类型：
--
作者：
Zhang, D;an;Zhang, Lida;Wang, Dapeng;Suo, Biao;Shi, Xianming
单核细胞增生李斯特菌是一种机会性食源性病原体，在全球范围内对健康造成严重危害。本研究建立了一种检测L.使用通过比较基因组方法鉴定的新的物种特异性靶序列（lmo 0754基因的区域），将内部扩增对照并入该PCR系统中。该方法仅能从L.李斯特菌属菌株，而不是来自其他李斯特菌属物种，以及一些非李斯特菌属物种。该方法的检出限为55拷贝/PCR。单核细胞增生基因组DNA。应用此PCR方法检测人工污染的牛奶样品，低水平的L。在选择性培养物富集（UVM 1）中孵育6-9小时后，检测到单核细胞增多症（1-10 cfu/mL牛奶）。
实际应用
通过比较基因组学方法鉴定了一个新的种特异性靶序列，为单核细胞增生李斯特菌的特异性检测提供了一个新的分子诊断标记。利用该靶标，建立了一种具有内扩增对照的PCR检测方法，该方法具有特异性、灵敏度和适用性。从人工污染的食物中提取单核细胞增多症。此外，这种比较基因组的方法也可以提供一个新的工具，挖掘独特的目标，为其他细菌的基因组序列的公共数据的增加。
Listeria monocytogenes is an opportunistic foodborne pathogen that is a serious health hazard worldwide. In this study, a polymerase chain reaction (PCR) assay was developed for the detection of L. monocytogenes using a novel species-specific target sequence (a region of lmo0754gene) identified by a comparative genomic approach. An internal amplification control was incorporated into this PCR system. The assay allowed amplification of a 331-bp fragment only from the genomic DNA of L. monocytogenes strains and not from other Listeria species, as well as some non-Listeria species. The detection limit of the PCR assay was 55 copies/PCR with L. monocytogenes genomic DNA. Applying this PCR assay to artificially contaminated milk samples, low levels of L. monocytogenes (1–10 cfu/mL of milk) were detected after 6–9 h incubation in selective culture enrichment (UVM1).
PRACTICAL APPLICATIONS
A novel species-specific target sequence was identified by a comparative genomic approach that offered a new molecular diagnostic marker for specific detection of Listeria monocytogenes. Using this target, a PCR assay with an internal amplification control was developed and shown to be specific, sensitive and applicable to the detection of L. monocytogenes from artificially contaminated foods. Moreover, this comparative genomic approach could also provide a new tool in mining unique targets for other bacteria with the increasing availability of public data of genome sequences.
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影响因子：
6.1
作者：
Alarcón, B;García-Cañas, V;Aznar, R
通讯作者：
Aznar, R
影响因子：
3.1
作者：
Chatterjee, SS;Hossain, H;Hain, T
通讯作者：
Hain, T
DOI：
10.1016/j.diagmicrobio.2007.06.011
发表时间：
2007-12
影响因子：
2.9
作者：
Zhanmin Liu;Xianming Shi;Feng Pan
通讯作者：
Zhanmin Liu;Xianming Shi;Feng Pan
DOI：
--
发表时间：
2001-10
期刊：
The new microbiologica
影响因子：
--
作者：
D. Pangallo;E. Kaclíková;T. Kuchta;Hana Drahovská
通讯作者：
D. Pangallo;E. Kaclíková;T. Kuchta;Hana Drahovská
影响因子：
12.6
作者：
Kim, Hyun-Joong;Park, Si-Hong;Kim, Hae-Yeong
通讯作者：
Kim, Hae-Yeong