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16S rRNA 测序鉴定1株条件致病性人苍白杆菌
引用本文:洪文艳,李曦,刘建伟,于德宪,谢晓波,刘金华,唐博恒.16S rRNA 测序鉴定1株条件致病性人苍白杆菌[J].国际检验医学杂志,2014,0(14):1819-1820.
作者姓名:洪文艳  李曦  刘建伟  于德宪  谢晓波  刘金华  唐博恒
作者单位:洪文艳 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 李曦 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 刘建伟 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 于德宪 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 谢晓波 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 刘金华 (广州军区疾病预防控制中心疾病监控科,广东广州,510507); 唐博恒 (广州军区疾病预防控制中心疾病监控科,广东广州,510507);
基金项目:艾滋病和病毒性肝炎等重大传染病防治”科技重大专项传染病监测技术平台项目(2009-ZX10004-205).
摘    要:目的:探索一种基于16S rRNA 基因的细菌快速鉴定方法,为临床未知病原菌的诊断及治疗提供科学依据。方法对临床患者的痰标本分离培养纯菌落,直接以菌液为模板,以通用引物 PCR 扩增未知菌的16S rRNA 基因片段,产物直接测序。将测序结果进行 BLAST 比对,根据序列同源性鉴定病原细菌。结果未知病原菌经本实验鉴定为人苍白杆菌,经 ABI 细菌快速鉴定板条检测,确认结果一致。结论该研究简化了临床标本未知病原菌分离培养鉴定的步骤,建立了一种利用16S rRNA 基因扩增快速鉴定病原菌的简便方法。

关 键 词:病原检测  细菌鉴定  直接PCR  16S  rRNA

Identifying 1 strain of conditional pathogenic Ochrobactrum by 16S rRNA gene sequencing
Hong Wenyan,Li Xi,Liu Janwei,Yu Dexian,Xie Xiaobo,Liu Jinhua,Tang Boheng.Identifying 1 strain of conditional pathogenic Ochrobactrum by 16S rRNA gene sequencing[J].International Journal of Laboratory Medicine,2014,0(14):1819-1820.
Authors:Hong Wenyan  Li Xi  Liu Janwei  Yu Dexian  Xie Xiaobo  Liu Jinhua  Tang Boheng
Institution:(Department of Disease Monitoring and Control ,Center of Disease Prevention and Control of Guangzhou Militerary Region Guangzhou Guangdong 510507, China)
Abstract:Objective To explore a rapid bacterial identifying method based on the 16S rRNA gene sequence analysis technology to provide the scientific basis for the diagnosis and treatment of unknown pathogenic bacteria.Methods The pure colonies were iso-lated and cultured directly from a clinical patient′s sputum sample.The colony as a template for PCR amplification with universal primers to amplify 16S rRNA gene fragments of unknown bacteria.The product of PCR was sequenced directly,then the sequence result was compared by using the BLAST of NCBI and the pathogen was identified based on the sequence homology.Results 1 strain of unknown pathogen was identified as ochrobactrum by this test and confirmed by ABI bacterial rapid identification sys-tem.Conclusion This study simplifies the isolation and identification procedures of unknown pathogen from the clinical samples and establishes a simple method for the rapid identification of pathogens by using 16S rRNA gene amplification.
Keywords:pathogen detection  bacterial identification  direct PCR  16S rRNA
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