Redondo Javier Jareño, Keller Peter M, Zbinden Reinhard, Wagner Karoline
Institute of Medical Microbiology, University of Zurich, Switzerland.
Institute of Medical Microbiology, University of Zurich, Switzerland.
Diagn Microbiol Infect Dis. 2018 Jan;90(1):1-6. doi: 10.1016/j.diagmicrobio.2017.09.014. Epub 2017 Sep 25.
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 97bp 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 256mg/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.
在本研究中，我们评估了市售的LightMix® RT-PCR检测法，用于检测幽门螺杆菌以及鉴定培养物和临床标本（胃活检组织和粪便）中的克拉霉素（CLR）耐药性。幽门螺杆菌LightMix® RT-PCR检测23S rRNA基因的一个97bp长的片段，并通过熔解曲线分析鉴定出导致CLR耐药的3种不同点突变。使用一组60株幽门螺杆菌菌株评估幽门螺杆菌LightMix® RT-PCR的性能，这些菌株表现出对CLR的表型敏感性或CLR耐药性（最低抑菌浓度为0.016至256mg/L）。我们发现，通过E-Test®进行的CLR耐药表型筛查与Lightmix® RT-PCR之间具有高度一致性（95%）。通过对23S rRNA基因进行测序验证了不一致的结果，测序结果始终证实了Lightmix® RT-PCR获得的结果。此外，通过Lightmix® RT-PCR在临床活检组织和粪便标本中检测到了幽门螺杆菌，并鉴定出了正确的幽门螺杆菌基因型。LightMix® RT-PCR是一种用于检测幽门螺杆菌和CLR耐药性的准确、灵敏且易于使用的检测方法，因此可以在任何诊断实验室中轻松实施。
