与肉汤微量稀释法相比,评估 MBT 脂质提取试剂盒在评估秋水仙碱耐药性方面的准确性。

Calvin Ka-Fung Lo, Gordon Ritchie, Jennifer Bilawka, Leah Gowland, Samuel D Chorlton, Willson Jang, Nancy Matic, Marc G Romney, Aleksandra Stefanovic, Christopher F Lowe
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引用次数: 0

摘要

肉汤微量稀释法(BMD)等大肠菌素耐药性检测方法对于临床实验室来说既耗时又耗力。MALDI Biotyper Sirius 系统(Bruker,Billerica,MA,USA)上的 MBT 脂质提取试剂盒利用脂质体分析来鉴定与秋水仙碱耐药性相关的特定细胞壁修饰。我们对 36 个革兰氏阴性分离物(非耐药 MIC ≤2 µg ml-1,耐药 MIC ≥4 µg ml-1)的 MBT 和 BMD(ComASP Colistin,Liofilchem)进行了比较。在评估 MBT 和 BMD 的分类一致性之前,对所有样本进行了两次 MBT 检测,并重复了不一致的结果。44.4%(16/36)的分离物通过 BMD 检测出对大肠菌素耐药。MBT Lipid Xtract 与 BMD 的吻合率为 80.6%(29/36),其中 5/7 的差异在重复检测后被纠正为吻合。大肠埃希菌分离物的一致性为 100%(n=16)。对两个不一致的肺炎克雷伯菌分离物完成了全基因组测序,确定了可乐定耐药性相关位点的变异(MIC 0.5 µg ml-1:arnC S30T、pmrB T246A、lapB N212T、lpxM S253G、crrB Q287K;MIC >16 µg ml-1:arnC S30T、pmrB R90insRN、pmrB T246A、pmrA E57G、lpxM S253G)。在临床实验室实施之前,需要对 MBT 进行进一步的评估,尤其是针对非大肠杆菌的评估。
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Evaluating the accuracy of the MBT Lipid Xtract Kit for assessing colistin resistance in comparison to broth microdilution.

Colistin resistance testing methods such as broth microdilution (BMD) are time-consuming and labour intensive for clinical laboratories. MBT Lipid Xtract Kit on MALDI Biotyper Sirius System (Bruker, Billerica, MA, USA) utilizes lipidomic analysis to identify specific cell wall modifications associated with colistin resistance. We compared MBT to BMD (ComASP Colistin, Liofilchem) across 36 Gram-negative isolates (non-resistant MIC ≤2 µg ml-1, resistant MIC ≥4 µg ml-1). All samples were tested twice on MBT with discrepant results repeated before assessing categorical agreement between MBT and BMD. 44.4% (16/36) of isolates were colistin resistant via BMD. MBT Lipid Xtract had 80.6% agreement (29/36) with BMD, with 5/7 discrepancies corrected to match upon repeat testing. There was 100% agreement for Escherichia coli isolates (n=16). The whole-genome sequencing was completed on the two discrepant Klebsiella pneumoniae isolates, with variants within colistin resistance-associated loci identified (MIC 0.5 µg ml-1: arnC S30T, pmrB T246A, lapB N212T, lpxM S253G, crrB Q287K and MIC >16 µg ml-1: arnC S30T, pmrB R90insRN, pmrB T246A, pmrA E57G, lpxM S253G). Further evaluation, particularly for non-E. coli, of MBT is required prior to implementation in clinical laboratories.

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