Frequent Suboptimal Thermocycler Ramp Rate Usage Negatively Impacts GenoType MTBDRsl VER 2.0 Performance for Second-Line Drug-Resistant Tuberculosis Diagnosis
Frequent Suboptimal Thermocycler Ramp Rate Usage Negatively Impacts GenoType MTBDRsl VER 2.0 Performance for Second-Line Drug-Resistant Tuberculosis Diagnosis
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DOI：
10.1016/j.jmoldx.2022.01.003
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发表时间：
2022-05-12
影响因子：
4.1
通讯作者：
Theron, Grant
中科院分区：
文献类型：
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作者：
Derendinger, Brigitta;de Vos, Margaretha;Theron, Grant
加强二线耐药结核病（TB）检测是一个优先事项。基因型MTBDRplus VER 2.0性能降低与非推荐的斜坡率使用（PCR周期之间的温度变化速度）；然而，斜坡率对基因型MTBDRsl VER 2.0 （MTBDRsl）性能的影响尚不清楚。在制造商推荐的（2.2℃/秒）或次优（4.0℃/秒）斜坡速率下，对52例Xpert MTB/RIF超阳性利福平耐药涂片阴性痰液和结核分枝杆菌稀释系列进行了测试。评估结核分枝杆菌-复合体- dna阳性，不确定，氟喹诺酮和二线注射剂-耐药准确性，带差异，以及单独的阅读器间变异性。13个重新调查的实验室中有5个（39%）没有使用制造商推荐的斜坡率。在痰液方面，2.2摄氏度/秒改善了不确定度，而4.0摄氏度/秒改善了不确定度（52中的0比51中的7，P = 0.006），错误的药物分类诊断呼叫（104中的0比102中的6，P = 0.013），错误的分级呼叫（1300中的0比1275中的54，P < 0.001）。同样，2.2℃/秒改善了有效结果[(52 / 52 vs 51 / 41; +21% (P = 0.001)]和频带呼叫阅读器间变异性[1300 / 34 (3%)vs 1300 / 52 (4%)]；P = 0.030]。在次优斜坡率下，由于野生型条带缺失而非突变型条带出现而产生假抗性和假敏感召唤，导致莫西沙星耐药水平从高到低的错误分类。次优斜坡速率导致MTBDRsl性能较差。实验室必须确保使用制造商推荐的斜坡速率。（分子诊断杂志，2022,24:494-502;https://doi.org/ 10.1016/j.jmoldx.2022.01.003）
Strengthening second-line drug-resistant tuberculosis (TB) detection is a priority. GenoType MTBDRplus VER 2.0 performance is reduced with non-recommended ramp rate usage (temperature change speed between PCR cycles); however, ramp rate's effect on GenoType MTBDRsl VER 2.0 (MTBDRsl) performance, is unknown. Fifty-two Xpert MTB/RIF Ultra-positive rifampicin-resistant smear-negative sputa and a Mycobacterium tuberculosis dilution series were tested at a manufacturer-recommended (2.2 degrees C/second) or suboptimal (4.0 degrees C/second) ramp rate. M. tuberculosis-complex-DNA positivity, indeterminates, fluoroquinolone- and second-line injectable-resistance accuracy, banding differences, and, separately, inter-reader variability were assessed. Five (39%) of 13 re-surveyed laboratories did not use the manufacturer-recommended ramp rate. On sputum, 2.2 degrees C/second improved indeterminates versus 4.0 degrees C/second (0 of 52 versus 7 of 51; P = 0.006), incorrect drug-class diagnostic calls (0 of 104 versus 6 of 102; P = 0.013), and incorrect banding calls (0 of 1300 versus 54 of 1275; P < 0.001). Similarly, 2.2 degrees C/second improved valid results [(52 of 52 versus 41 of 51; +21% (P = 0.001)] and banding call inter-reader variability [34 of 1300 (3%) versus 52 of 1300 (4%); P = 0.030]. At the suboptimal ramp rate, false-resistance and false-susceptible calls resulted from wild-type band absence rather than mutant band appearance, resulting in misclassification of moxifloxacin resistance level from high-tolow. Suboptimal ramp rate contributes to poor MTBDRsl performance. Laboratories must ensure that the manufacturer-recommended ramp rate is used. (J Mol Diagn 2022, 24: 494-502; https://doi.org/ 10.1016/j.jmoldx.2022.01.003)