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检测菌毛因子和分枝杆菌菌落的宏观评估:固体培养基中用于疑似结核分枝杆菌复合群鉴定的一种高效联合筛选试验。

Cord factor detection and macroscopic evaluation of mycobacterial colonies: an efficient combined screening test for the presumptive identification of Mycobacterium tuberculosis complex on solid media.

机构信息

Instituto Adolfo Lutz, São Paulo, Brazil.

出版信息

J Bras Pneumol. 2009 Dec;35(12):1212-6. doi: 10.1590/s1806-37132009001200008.

Abstract

OBJECTIVE

The rapid differentiation between Mycobacterium tuberculosis and nontuberculous mycobacteria is fundamental for patients co-infected with tuberculosis and HIV. To that end, we use two methods in our laboratory: detection of cord factor and PCR-restriction enzyme analysis (PRA). The objective of this study was to evaluate the accuracy of a screening test on solid medium as a rapid method for the presumptive identification of M. tuberculosis complex, considering costs and turnover time.

METHODS

A total of 152 strains were submitted to a combined screening test, consisting of the detection of cord factor under microscopy (Ziehl-Neelsen staining) and evaluation of the macroscopic aspect of colonies, as well as to PRA, which was used as the gold standard. Costs were estimated by calculating the price of all of the materials needed for each test.

RESULTS

The overall accuracy of cord factor detection alone was 95.4% (95% CI: 90.7-98.1%), and that of the combined screening test was 99.3% (95% CI: 96.4-100%). Cord factor detection costs US$ 0.25, whereas the PRA costs US$ 7.00. Results from cord factor detection are ready in 2 days, whereas PRA requires 4 days to yield results.

CONCLUSIONS

The presumptive identification of M. tuberculosis using the macroscopic evaluation of colonies combined with cord factor detection under microscopy is a simple, rapid and inexpensive test. We recommend the combined screening test to rapidly identify M. tuberculosis in resource-poor settings and in less well-equipped laboratories while awaiting a definite identification by molecular or biochemical methods.

摘要

目的

结核分枝杆菌和非结核分枝杆菌的快速区分对于合并结核分枝杆菌和 HIV 感染的患者至关重要。为此,我们在实验室中使用两种方法:检测 cord 因子和 PCR-限制性内切酶分析(PRA)。本研究旨在评估固体培养基上的筛选试验作为快速方法对结核分枝杆菌复合群进行初步鉴定的准确性,同时考虑成本和周转时间。

方法

共对 152 株菌株进行联合筛选试验,包括显微镜下检测 cord 因子(Ziehl-Neelsen 染色)和评估菌落的宏观外观,以及作为金标准的 PRA。通过计算每个测试所需的所有材料的价格来估算成本。

结果

单独检测 cord 因子的总准确率为 95.4%(95%置信区间:90.7-98.1%),联合筛选试验的准确率为 99.3%(95%置信区间:96.4-100%)。cord 因子检测的成本为 0.25 美元,而 PRA 的成本为 7.00 美元。cord 因子检测的结果可在 2 天内得出,而 PRA 需要 4 天才能得出结果。

结论

使用显微镜下的菌落宏观评估结合 cord 因子检测来初步鉴定结核分枝杆菌是一种简单、快速且廉价的方法。我们建议在资源匮乏的环境中和设备较差的实验室中使用联合筛选试验,以快速鉴定结核分枝杆菌,同时等待分子或生化方法进行明确鉴定。

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