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使用快速刃天青微量滴定板法对结核分枝杆菌进行吡嗪酰胺药敏试验。

Pyrazinamide susceptibility testing in Mycobacterium tuberculosis using the fast resazurin microtiter assay plate.

作者信息

Pina R Z, Caleffi-Ferracioli K R, Campanerut-Sá P A Z, Ghiraldi-Lopez L D, Pavan F R, Siqueira V L D, Scodro R B L, Cardoso R F

机构信息

Post-Graduate Programme in Health Sciences, State University of Maringá, Maringá, Brazil.

Department of Clinical Analyses and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.

出版信息

Int J Tuberc Lung Dis. 2016 Nov;20(11):1535-1538. doi: 10.5588/ijtld.16.0304.

Abstract

SETTING

Department of Clinical Analysis and Biomedicine, State University of Maringa, Maringa, PR, Brazil.

OBJECTIVE

To evaluate the performance of the resazurin microtiter assay (REMA) plate at pH 5.5 in detecting Mycobacterium tuberculosis susceptibility to pyrazinamide (PZA).

DESIGN

The minimal inhibitory concentration (MIC) of PZA in M. tuberculosis HRv and M. bovis AN5 reference strains and in 34 clinical M. tuberculosis isolates (26 PZA-susceptible and eight PZA-resistant) was determined using REMA at pH 5.5 and compared to REMA at pH 6.0.

RESULTS

REMA at pH 5.5 was helpful in discriminating PZA-susceptible from resistant M. tuberculosis isolates when ⩿50 μg/ml PZA was considered as the cut-off for PZA susceptibility. Furthermore, it provided results in 8 days. However, two PZA-resistant isolates failed to grow at pH 5.5.

CONCLUSION

As the REMA method is rapid, inexpensive, easy to perform and read, it would be of great usefulness in low-income countries for detecting PZA-resistant M. tuberculosis. REMA at pH 5.6-5.9 should be evaluated on an extended panel of clinical M. tuberculosis isolates with a greater range of MIC values in different laboratories for a better understanding of its utility in differentiating PZA-resistant from PZA-susceptible isolates.

摘要

背景

巴西巴拉那州马林加市州立大学临床分析与生物医学系。

目的

评估刃天青微量滴定法(REMA)平板在pH 5.5条件下检测结核分枝杆菌对吡嗪酰胺(PZA)敏感性的性能。

设计

使用pH 5.5的REMA法测定结核分枝杆菌H37Rv和牛分枝杆菌AN5参考菌株以及34株临床结核分枝杆菌分离株(26株对PZA敏感,8株对PZA耐药)中PZA的最低抑菌浓度(MIC),并与pH 6.0的REMA法进行比较。

结果

当将PZA浓度⩿50 μg/ml作为PZA敏感性的临界值时,pH 5.5的REMA法有助于区分对PZA敏感和耐药的结核分枝杆菌分离株。此外,该方法在8天内得出结果。然而,有两株对PZA耐药的分离株在pH 5.5条件下无法生长。

结论

由于REMA方法快速、廉价、易于操作和判读,在低收入国家检测耐PZA的结核分枝杆菌时将非常有用。应在不同实验室中,对更多具有更广泛MIC值范围的临床结核分枝杆菌分离株进行评估,以更好地了解pH 5.6 - 5.9的REMA法在区分耐PZA和对PZA敏感分离株方面的效用。

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