16S rRNA序列分析技术对临床标本中疑难细菌的鉴定  

Application of 16S rRNA gene sequence analysis for identification of clinical unidentifiable bacteria

作　　者：叶乃芳 凌华志[1] 黄颖[1] 沈继录[1] 徐元宏[1] 王中新[1] 

机构地区：[1]安徽医科大学第一附属医院检验科,合肥230022

出　　处：《安徽医科大学学报》2015年第7期1000-1003,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校省级自然科学研究项目(编号:KJ2010A174)

摘　　要：目的应用16S rRNA序列分析技术鉴定临床少见或疑难细菌,提高细菌鉴定的准确性。方法收集临床少见或疑难细菌44株,提取DNA,采用通用引物进行PCR扩增及目的片段基因测序,并将测序结果在核酸数据库中比对分析以确定菌种。结果 44株临床少见或疑难细菌均扩增到16S rRNA目的基因片段并成功测序,40株(90.9%)鉴定到"种",4株(9.1%)鉴定到"属";其中常规方法与测序方法在"属"水平一致的有34株(77.3%),不一致的有10株(22.7%)。结论 16S rRNA序列分析技术可以准确、快速地鉴定临床少见或疑难细菌,可以作为临床细菌鉴定的重要方法之一。Objective To apply an approach based on 16 S rRNA gene sequences analysis for identification of clini-cal rare and unidentifiable bacteria in order to improve the accuracy of clinical diagnosis. Methods A total of 44 clinical isolates were collected,the bacteria DNA was extracted followed by PCR amplification for 16S rRNA gene with a universal primer. The PCR products were sequenced . The confirmed sequences were compared to the refer-ence sequences obtained from GenBank and RDP databases. Results All of the 44 clinical isolates obtained the target genes successfully. 40 strains （90. 9%） were specified to the species level and 4 strains （9. 1%） were specified to the genus level. 77. 3%（ 34/44 ） of sequencing results were concordant with phenotypic results, 22. 7%（10/44） of sequencing results were not concordant with phenotypic results. Conclusion 16S rRNA gene sequence analysis can be accurate, rapid identification of clinical rare bacteria and bacteria with unusual phenotyp-ic profiles.

关 键 词：16S RRNA基因 序列分析 细菌 鉴定 

分 类 号：R446.5[医药卫生—诊断学]