摘要
目的用焦磷酸测序技术鉴定经VITEK 2 Compact系统鉴定准确率低或无法鉴定的46株细菌,对两种方法在少见菌的鉴定能力进行评价。方法选择经VITEK 2 Compact系统鉴定准确率低或无法鉴定的46株细菌(包含革兰阳性球菌、革兰阳性杆菌、革兰阴性球菌及革兰阴性杆菌),提取DNA模板,做焦磷酸测序技术分析;同步进行Sanger法测序并以其结果作为准确鉴定的"金标准"。结果焦磷酸测序技术在4 h内完成46株细菌的鉴定。焦磷酸测序技术鉴定正确率为71.74%(33/46),VITEK 2 Compact系统鉴定正确率为43.48%(20/46)。46株菌中,两种方法鉴定均鉴定正确的菌株19株;焦磷酸测序技术鉴定正确、VITEK 2 Compact系统鉴定准确率低、未鉴定或鉴定错误的菌株14株;VITEK2 Compact系统鉴定正确、焦磷酸测序技术鉴定错误的菌株1株;两种方法均鉴定错误的菌株12株。结论对于微生物实验室利用传统方法无法准确鉴定的细菌,利用焦磷酸测序技术鉴定具有准确、快速及简便的特点,可适用于临床少见病原菌的鉴定。
Objective To identify 46 bacteria by pyrosequencing system which with low reliability or unable to be identified by VITEK 2 Compact system, and evaluate the rare bacteria identification ability of the two methods. Methods Forty-six bacteria with low reliability or unable to be identified by VITEK 2 Compact system were selected（included gram positive cocci, gram positive bacillus, gram negative cocci and gram negative bacillus）, the DNA was extracted and analyzed by pyrosequenc- ing. The Sanger sequencing was performed at the same time and results were taken as ＂gold standard＂. Results The pyrose- quening identified 46 bacteria in 4 hours; The identification accuracy of pyrosequening was 71.74 %（33/46） and that of VITEK 2 Compact was 43.48 %（20/46）. In 46 bacteria, 19 were identified correctly by 2 methods and 14 were correctly identified by pyrosequeneing, while VITEK 2 Compact test showed low resolution, unable to be identified or error results. One of the bacte- ria was identified correctly by VITEK 2 Compact but error result by pyrosequencing, and 12 of the bacteria were error identi- fied by both methods. Conclusion It is demonstrated that bacteria cannot be identified accurately by traditional identification method in clinical microorganism laboratory, identification with pyrosequencing shows accuracy, convenience and high speed. It can be applied to identify clinical rare pathogen.
出处
《生物医学工程与临床》
CAS
2016年第6期626-630,共5页
Biomedical Engineering and Clinical Medicine
基金
济南军区总医院院长基金资助项目(2014QN08)
