Gascoyne-Binzi D M, Barlow R E, Frothingham R, Robinson G, Collyns T A, Gelletlie R, Hawkey P M
Department of Microbiology, The General Infirmary, Leeds LS1 3EX, United Kingdom.
J Clin Microbiol. 2001 Jan;39(1):69-74. doi: 10.1128/JCM.39.1.69-74.2001.
Compared with solid media, broth-based mycobacterial culture systems have increased sensitivity but also have higher false-positive rates due to cross-contamination. Systematic strain typing is rarely undertaken because the techniques are technically demanding and the data are difficult to organize. Variable number tandem repeat (VNTR) analysis by PCR is rapid and reproducible. The digital profile is easily manipulated in a database. We undertook a retrospective study of Mycobacterium tuberculosis isolates collected over an 18-month period following the introduction of the BACTEC MGIT 960 system. VNTR allele profiles were determined with early positive broth cultures and entered into a database with the specimen processing date and other specimen data. We found 36 distinct VNTR profiles in cultures from 144 patients. Three common VNTR profiles accounted for 45% of true-positive cases. By combining VNTR results with specimen data, we identified nine cross-contamination incidents, six of which were previously unsuspected. These nine incidents resulted in 34 false-positive cultures for 29 patients. False-positive cultures were identified for three patients who had previously been culture positive for tuberculosis and were receiving treatment. Identification of cross-contamination incidents requires careful documentation of specimen data and good communication between clinical and laboratory staff. Automated broth culture systems should be supplemented with molecular analysis to identify cross-contamination events. VNTR analysis is reproducible and provides timely results when applied to early positive broth cultures. This method should ensure that patients are not placed on unnecessary tuberculosis therapy or that cases are not falsely identified as treatment failures. In addition, areas where existing procedures may be improved can be identified.
与固体培养基相比,基于肉汤的分枝杆菌培养系统提高了敏感性,但由于交叉污染也有更高的假阳性率。由于技术要求高且数据难以整理,很少进行系统的菌株分型。通过聚合酶链反应(PCR)进行可变数目串联重复序列(VNTR)分析快速且可重复。数字图谱在数据库中易于处理。我们对引入BACTEC MGIT 960系统后18个月期间收集的结核分枝杆菌分离株进行了一项回顾性研究。用早期阳性肉汤培养物确定VNTR等位基因图谱,并将其与标本处理日期和其他标本数据一起输入数据库。我们在144例患者的培养物中发现了36种不同的VNTR图谱。三种常见的VNTR图谱占真阳性病例的45%。通过将VNTR结果与标本数据相结合,我们识别出9起交叉污染事件,其中6起以前未被怀疑。这9起事件导致29例患者出现34例假阳性培养结果。在3例先前结核培养阳性且正在接受治疗的患者中发现了假阳性培养结果。识别交叉污染事件需要仔细记录标本数据以及临床和实验室工作人员之间的良好沟通。自动化肉汤培养系统应辅以分子分析以识别交叉污染事件。VNTR分析可重复,应用于早期阳性肉汤培养物时能提供及时的结果。这种方法应确保患者不会接受不必要的结核病治疗,或病例不会被错误地判定为治疗失败。此外,还可以确定现有程序可能需要改进的方面。