广西市售婴儿食品中克罗诺菌分离株基因特征及进化分类鉴定
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(广西壮族自治区疾病预防控制中心,广西 南宁 530028)

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王红 女 副主任技师 研究方向为微生物检验及分子生物学 E-mail:gxcdcw@126.com通信作者:┣┣(中)通信作者┫┫黄彦 男 副主任技师 研究方向为微生物检验及分子生物学 E-mail:huangyangx@163.com

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广西自然科学基金(2012GXNSFBA053125);广西科学研究与技术开发计划(桂科攻1298004-3)


Identification,classification,phenotypic and genetic characterization of Cronobacter spp. isolated from infant food
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(Guangxi Zhuang Autonomous Region Center for Disease Control and Prevention, Guangxi Nanning 530028,China)

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    摘要:

    分析广西市售婴儿食品中的克罗诺菌分离株的种类、表型和基因特性。方法 将保存的试验菌株进行复苏,通过API 20E生化条和ompA基因扩增检测进行初步鉴定,扩增16S rRNA基因后进行测序,将获得的全16S rRNA基因序列在GeneBank数据库上比对,构建进化树,确认是否为克罗诺菌。将试验菌接种于显色平板观察其表型和黄色素的产生情况,通过手工生化进行分型,确定其种属。结果 9株分离菌确定为克罗诺菌,有5种生化型,属于4个克罗诺种。检出Cronobacter sakazakii 6株菌,C.malonaticus、C.muytjensii和C.dublinensis各有1株;7株菌符合 API 20E鉴定结果,9株分离菌均可检测到ompA基因。结论 广西市售婴儿食品中克罗诺菌的污染以C.sakazakii为主,生化表型、种属及基因型具有多样性;鉴定时仅以单一的生化或其他表型作为判别依据存在很大的局限,需辅以分子生物学手段加以鉴别。

    Abstract:

    To investigate the subspecies, phenotypic and genetic characterization of Cronobacter isdates from infant food in 2010.Methods All isolates were reactivated, then suspectable stains were identified preliminarily by two methods, PCR amplifying ompA gene and API 20E. To confirm these stains, full 16S rRNA gene sequences were obtained, and made BLASTN on GeneBank database. The identified strains were streaked onto DFI chromogenic agar and TSA to observe phenotype and yellow pigment production. Biochemical tests were conducted and assigned to biogroups to determine their species.Results Nine strains were identified as Cronobacter spp. These strains were assigned to 5 biogroups, and belonged to 4 species. The majority of strains were Cronobacter sakazakii, and three strains was identified as C.malonaticus, C.muytjensii and C.dublinensis respectively. 7 strains were identified as Cronobacter by API 20E, yet ompA gene amplification of all strains was positive.Conclusion Cronobacter isolated from infant food were diversed in biochemical phenotype and species. There was a risk of Cronobacter in infant food from supermarkets. There was limitation to identify the bacteria only by biochemical phenotype, and supplementary molecular biology methods was necessary.

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王红,黄彦,周艳,韦程媛,李秀桂,谭冬梅,诸葛石养,孙贵娟.广西市售婴儿食品中克罗诺菌分离株基因特征及进化分类鉴定[J].中国食品卫生杂志,2016,28(1):6-10.

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  • 收稿日期:2015-09-09
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  • 在线发布日期: 2016-02-19
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