贵州医科大学学报

2014, v.39;No.171(06) 805-808

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新生儿肠炎粪肠球菌16S rRNA鉴定分析
Identification of 16S rRNA in Neonatal Enteritis Caused by Enterococcus faecalis

康颖倩;牟丽丽;周晓明;王颜颜;刘涛华;金方;吕倩;
KANG Yingqian;MU Lili;ZHOU Xiaoming;WANG Yanyan;LIU Taohua;JIN Fang;LV Qian;Department of Microbiology,Guiyang Medical University;Guiyang Maternal and Child Health Hospital;

摘要(Abstract):

目的:分离和鉴定新生儿肠炎患儿便血中的病原菌,结合临床诊断,分析新生儿肠炎病因。方法:对2例新生儿肠炎病例的便血标本进行研究,(1)直接涂片镜检观察鉴定;(2)菌株纯化筛选;(3)采用CTAB提取法提取菌株的DNA,并通过PCR扩增菌株16S rRNA序列,将所获得的序列与Gen Bank数据库进行BLAST比对;(4)采用Kirby-Bauey纸片扩散法进行药物敏感试验。结果:(1)便血标本共分离纯化2株可疑菌株;(2)通过16S rRNA序列与Gen Bank数据库进行BLAST比对,两株菌株的序列与粪肠球菌的相似度达(99%);(3)分离的粪肠球菌对氨苄西林敏感性较高。结论:新生儿肠炎是由粪肠球菌感染引起,标本直接镜检与药敏试验有利于早期诊断及抗生素的合理应用。
Objective: To isolate and identify the pathogenic microbes from the bloody stool of newborn infant with neonatal enteritis,and to analyze the causes of neonatal enteritis with clinical diagnosis and therapy. Methods: Bloody stool of two newborn infants with neonatal enteritis was studied,( 1)Direct smear microscopy was applied for primary identification;( 2) The microbes were purified and screened;( 3) Total DNA of E. faecalis strains was extracted with CTAB rapid extraction protocol and the 16 S rRNA sequence data were amplified,obtained sequences were compared with Gen Bank database by BLAST;( 4) The drug susceptibility was tested by Kirby-Bauey method towards the isolated strains in vitro. Results:( 1) Two E. faecalis strains were isolated and purified form stool of newborn infant;( 2) 16 S rRNA sequence were compared with Gen Bank database by BLAST,sequence of two strains were similar with E. faecali strains( 99%);( 3) Drug susceptibility results showed that the clinical E. faecalis strains were sensitive to ampicillin and penicillin; Conclusions: Etiological analysis showed E. faecalis leaded to neonatal enteritis. Combined with molecular identification methods,early direct smear microscopy and drug susceptibility test in clinical labs can reduce the mortality rate of newborn infants and prevent the outbreak of infectious diseases effectively.

关键词(KeyWords): 肠炎;粪肠球菌;婴儿,新生;鉴定分析
neonatal enteritis;Enterococcus faecalis;infant,newborn;identification and analysis

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基金项目(Foundation): 国家自然基金地方项目(31060006,31260029);; 贵州省社会发展科技攻关项目[黔科合SY字(2011)3017号];; 贵阳市科技局社会发展与民生计划[筑科合同(2011103)16号]

作者(Author): 康颖倩;牟丽丽;周晓明;王颜颜;刘涛华;金方;吕倩;
KANG Yingqian;MU Lili;ZHOU Xiaoming;WANG Yanyan;LIU Taohua;JIN Fang;LV Qian;Department of Microbiology,Guiyang Medical University;Guiyang Maternal and Child Health Hospital;

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