Validating a 14-Drug Microtiter Plate Containing Bedaquiline and Delamanid for Large-Scale Research Susceptibility Testing of Mycobacterium tuberculosis.
Validating a 14-Drug Microtiter Plate Containing Bedaquiline and Delamanid for Large-Scale Research Susceptibility Testing of Mycobacterium tuberculosis.
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DOI：
10.1128/aac.00344-18
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发表时间：
2018-09
影响因子：
4.9
通讯作者：
CRyPTIC Consortiumfor the CRyPTIC Consortiumfor the CRyPTIC Consortium
中科院分区：
文献类型：
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作者：
Rancoita PMV;Cugnata F;Gibertoni Cruz AL;Borroni E;Hoosdally SJ;Walker TM;Grazian C;Davies TJ;Peto TEA;Crook DW;Fowler PW;Cirillo DM;CRyPTIC Consortiumfor the CRyPTIC Consortiumfor the CRyPTIC Consortium
UKMYC 5板是由CRyPTIC联盟（结核病综合耐药预测：国际联盟）设计的96孔微量滴定板，能够测量14种不同抗结核（抗TB）化合物对> 30，000种临床结核分枝杆菌分离株的MIC。与UKMYC 5平板所基于的MYCOTB平板不同，UKMYC 5平板包括两种新化合物（贝达喹啉和德拉马尼）和两种改性化合物（氯法齐明和利奈唑胺）。UKMYC 5板是由CRyPTIC联盟（结核病综合耐药预测：国际联盟）设计的96孔微量滴定板，能够测量14种不同抗结核（抗TB）化合物对> 30，000种临床结核分枝杆菌分离株的MIC。与UKMYC 5平板所基于的MYCOTB平板不同，UKMYC 5平板包括两种新化合物（贝达喹啉和德拉马尼）和两种改性化合物（氯法齐明和利奈唑胺）。UKMYC 5平板由四大洲的七个实验室使用一组19种室间质量评估（EQA）菌株（包括H37 Rv）进行检测。为了评估阅读方法和孵育时间的最佳组合，在孵育7、10、14和21天后，由两名读数员使用三种方法（镜箱、显微镜和Vizion数字观察系统）测量每个平板的MIC。此外，对所有EQA菌株进行全基因组测序，并通过7 H10/7 H11琼脂比例法（APM）和使用MGIT 960分枝杆菌生长指示管进行表型表征。我们得出结论，培养14天后，使用Vizion系统可最佳读取UKMYC 5平板，读数员间一致性为97.9%，实验室内和实验室间重现性分别为95.6%和93.1%。镜盒也有类似的再现性。通过APM、MGIT 960或存在已知赋予耐药性的突变分类为耐药的菌株与分类为敏感的菌株相比，一致显示MIC升高。最后，UKMYC 5平板记录了一种菌株的中间MIC，APM测得的MIC接近应用的临界浓度，提供了早期证据，证明UKMYC 5平板可以定量测量由于特定遗传变异引起的抗结核化合物耐药性的程度。
The UKMYC5 plate is a 96-well microtiter plate designed by the CRyPTIC Consortium (Comprehensive Resistance Prediction for Tuberculosis: an International Consortium) to enable the measurement of MICs of 14 different antituberculosis (anti-TB) compounds for >30,000 clinical Mycobacterium tuberculosis isolates. Unlike the MYCOTB plate, on which the UKMYC5 plate is based, the UKMYC5 plate includes two new (bedaquiline and delamanid) and two repurposed (clofazimine and linezolid) compounds. The UKMYC5 plate is a 96-well microtiter plate designed by the CRyPTIC Consortium (Comprehensive Resistance Prediction for Tuberculosis: an International Consortium) to enable the measurement of MICs of 14 different antituberculosis (anti-TB) compounds for >30,000 clinical Mycobacterium tuberculosis isolates. Unlike the MYCOTB plate, on which the UKMYC5 plate is based, the UKMYC5 plate includes two new (bedaquiline and delamanid) and two repurposed (clofazimine and linezolid) compounds. UKMYC5 plates were tested by seven laboratories on four continents by use of a panel of 19 external quality assessment (EQA) strains, including H37Rv. To assess the optimal combination of reading method and incubation time, MICs were measured from each plate by two readers, using three methods (mirrored box, microscope, and Vizion digital viewing system), after 7, 10, 14, and 21 days of incubation. In addition, all EQA strains were subjected to whole-genome sequencing and phenotypically characterized by the 7H10/7H11 agar proportion method (APM) and by use of MGIT960 mycobacterial growth indicator tubes. We concluded that the UKMYC5 plate is optimally read using the Vizion system after 14 days of incubation, achieving an interreader agreement of 97.9% and intra- and interlaboratory reproducibility rates of 95.6% and 93.1%, respectively. The mirrored box had a similar reproducibility. Strains classified as resistant by APM, MGIT960, or the presence of mutations known to confer resistance consistently showed elevated MICs compared to those for strains classified as susceptible. Finally, the UKMYC5 plate records intermediate MICs for one strain for which the APM measured MICs close to the applied critical concentration, providing early evidence that the UKMYC5 plate can quantitatively measure the magnitude of resistance to anti-TB compounds that is due to specific genetic variation.