临床微生物实验室交叉污染警报的优化分子解决方案
Research articleOptimized molecular resolution of cross-contamination alerts in clinical mycobacteriology laboratories
临床微生物实验室交叉污染警报的优化分子解决方案
Ana Martin , Marta Herranz , Miguel Martinez Lirola , Rosa Fernandez Fernandez , Indal-tb Group , Emilio Bouzaand Dario Garcia de Viedma
BMC Microbiology 2008, 8:30doi:10.1186/1471-2180-8-30
Published: 14 February 2008
Abstract (provisional)
Background
The phenomenon of misdiagnosing tuberculosis by laboratory cross-contamination when culturing Mycobacterium tuberculosis (MTB) has been widely reported and it has an obvious clinical, therapeutic and social impact. The final confirmation of a cross-contamination event requires the molecular identification of the same MTB strain cultured from both the potential source of the contamination and from the false-positive candidate. The molecular tool usually applied in this context is IS6110-RFLP which takes a long time to provide an answer, usually longer than is acceptable for microbiologists and clinicians to make decisions. Our purpose in this study is to evaluate a novel PCR-based method, MIRU-VNTR as an alternative to assure a rapid and optimized analysis of cross-contamination alerts.
Results
MIRU-VNTR was prospectively compared with IS6110-RFLP for clarifying 19 alerts of false positivity from other laboratories. MIRU-VNTR highly correlated with IS6110-RFLP, reduced the response time by 27 days and clarified six alerts unresolved by RFLP. Additionally, MIRU-VNTR revealed complex situations such as contamination events involving polyclonal isolates and a false-positive case due to the simultaneous cross-contamination from two independent sources.
Conclusions
Unlike standard RFLP-based genotyping, MIRU-VNTR i) could help reduce the impact of a false positive diagnosis of TB, ii) increased the number of events that could be solved and iii) revealed the complexity of some cross-contamination events that could not be dissected by IS6110-RFLP.
Background(背景)
The phenomenon of misdiagnosing tuberculosis by laboratory cross-contamination when culturing Mycobacterium tuberculosis (MTB) has been widely reported and it has an obvious clinical, therapeutic and social impact. (由于实验室的交叉污染而大量的结核分支杆菌的假阳性报告使得实验室对结核的误诊,已经明显影响了临床诊断和治疗。)The final confirmation of a cross-contamination event requires the molecular identification of the same MTB strain cultured from both the potential source of the contamination and from the false-positive candidate. (交叉污染的最终确认依靠对来自潜在污染源和假阳性标本相同的结核分支杆菌进行分子鉴别。)The molecular tool usually applied in this context is IS6110-RFLP which takes a long time to provide an answer, usually longer than is acceptable for microbiologists and clinicians to make decisions.(通常使用的分子手段IS6110-RFLP需要很长时间才能得出结果,长时间的等待常常让微生物学家和临床医生无法接受。) Our purpose in this study is to evaluate a novel PCR-based method, MIRU-VNTR as an alternative to assure a rapid and optimized analysis of cross-contamination alerts.(我们的研究在于评价一种新的基于pcr的方法,MIRU-VNT,作为一种可选的手段,确保快速准确分析实验室的交叉污染警报。)
Results(结果)
MIRU-VNTR was prospectively compared with IS6110-RFLP for clarifying 19 alerts of false positivity from other laboratories. (MIRU-VNTR和IS6110-RFLP对19起假阳性警报进行分析比较。)MIRU-VNTR highly correlated with IS6110-RFLP, reduced the response time by 27 days and clarified six alerts unresolved by RFLP. (MIRU-VNTR与IS6110-RFLP高度相关,但反应时间减少了27天,且鉴别出了6起RFLP未能解决的警报。)Additionally, MIRU-VNTR revealed complex situations such as contamination events involving polyclonal isolates and a false-positive case due to the simultaneous cross-contamination from two independent sources.(另外,MIRU-VNTR能够反映出复杂状况,如多克隆株的鉴别,一起同时两处独立来源的交叉污染导致的假阳性事件。)
Conclusions(结论)
Unlike standard RFLP-based genotyping, MIRU-VNTR i) could help reduce the impact of a false positive diagnosis of TB,(不同于基于RFLP的基因分型,MIRU-VNTR能够帮助减少TB假阳性诊断的影响。) ii) increased the number of events that could be solved (增加了解决事件的数量)and iii) revealed the complexity of some cross-contamination events that could not be dissected by IS6110-RFLP.(解决复杂交叉污染事件,这些往往无法被IS6110-RFLP所处理。) 能不能介绍一下MIRU-VNTR(Mycobacterial-interspersedrepetitive-units-Variable-number-tandem-repeats)? 下载了好资料和微生物老师一起好好分享。。。真诚感谢老师的无私。。。问好。
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