摘要

在这项研究中，我们评估了商业上可用的LightMix®RT-PCR检测方法，用于幽门螺杆菌检测以及培养和临床标本（胃活检和大便）中克拉霉素（CLR）耐药性的鉴定。幽门螺旋杆菌LightMix®RT-PCR检测23S rRNA基因的97bp长的片段，并通过解链曲线分析鉴定3个不同的赋予CLR抗性的点突变。使用一组显示表型CLR敏感性或CLR抗性（最小抑制浓度为0.016至256mg / L）的60个幽门螺杆菌菌株评估幽门螺旋杆菌LightMix RT-PCR的性能。我们发现通过E-Test®和Lightmix®RT-PCR对表型CLR耐药筛查的高度一致性（95％）。通过23S rRNA基因的测序验证了不一致的结果，所述基因总是证实通过Lightmix RT-PCR获得的结果。此外，通过Lightmix RT-PCR在临床活检和粪便标本中检测到幽门螺旋杆菌，其鉴定了正确的幽门螺杆菌基因型。 LightMix®RT-PCR是一种准确，灵敏且易于使用的幽门螺杆菌和CLR耐药检测试剂，因此可以在任何诊断实验室中轻松实施。

In this study we evaluated the commercially available LightMix® RT-PCR assay for Helicobacter pylori detection and identification of clarithromycin (CLR) resistance in culture and clinical specimens (gastric biopsies and stool). The H. pylori LightMix® RT-PCR detects a 97 bp long fragment of the 23S rRNA gene and allows the identification of 3 distinct point mutations conferring CLR resistance via melting curve analysis. The performance of the H. pylori LightMix® RT-PCR was evaluated using a set of 60 H. pylori strains showing phenotypical CLR susceptibility or CLR resistance (Minimum inhibitory concentrations from 0.016 to 256 mg/L). We found high concordance (95%) between phenotypical CLR resistance screening by E-Test® and the Lightmix® RT-PCR. Discrepant results were verified by sequencing of the 23S rRNA gene that always confirmed the results obtained by Lightmix® RT-PCR. Furthermore, H. pylori was detected in clinical biopsy and stool specimens by Lightmix® RT-PCR that identified the correct H. pylori genotype. The LightMix® RT-PCR is an accurate, sensitive and easy to use test for H. pylori and CLR resistance detection and can therefore be readily implemented in any diagnostic laboratory.

http://www.dmidjournal.com/article/S0732-8893(17)30305-X/abstract