Kuang Zupeng, Huang Huishu, Chen Ling, Shang Yanyan, Huang Shixuan, Liu Jun, Chen Jianhui, Xie Xinqiang, Chen Moutong, Wu Lei, Gao He, Zhao Hui, Li Ying, Wu Qingping
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.
National Health Commission Science and Technology Innovation Platform for Nutrition and Safety of Microbial Food, Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China.
Antibiotics (Basel). 2024 Oct 16;13(10):975. doi: 10.3390/antibiotics13100975.
The issue of () resistance to clarithromycin (CLR) has consistently posed challenges for clinical treatment. Hence, a rapid susceptibility testing (AST) method urgently needs to be developed. In the present study, 35 isolates of were isolated from 203 gastritis patients of the Guangzhou cohort, and the antimicrobial resistance phenotypes were associated with their genomes to analyze the relevant mutations. Based on these mutations, a rapid detection system utilizing high-resolution melting (HRM) curve analysis was designed and verified by the Shenzhen cohort, which consisted of 38 strains. Genomic analysis identified the mutation of the 2143 allele from A to G (A2143G) of as the most relevant mutation with CLR resistance ( < 0.01). In the HRM system, the wild-type showed a melting temperature (Tm) of 79.28 ± 0.01 °C, while the mutant type exhibited a Tm of 79.96 ± 0.01 °C. These differences enabled a rapid distinction between two types of ( < 0.01). Verification examinations showed that this system could detect target DNA as low as 0.005 ng/μL in samples without being affected by other gastric microorganisms. The method also showed a good performance in the Shenzhen validation cohort, with 81.58% accuracy, and 100% specificity. We have developed an HRM system that can accurately and quickly detect CLR resistance in . This method can be directly used for the detection of gastric microbiota samples and provides a new benchmark for the simple detection of resistance.
（）对克拉霉素（CLR）的耐药性问题一直给临床治疗带来挑战。因此，迫切需要开发一种快速药敏试验（AST）方法。在本研究中，从广州队列的203例胃炎患者中分离出35株（）菌株，并将其抗菌耐药表型与其基因组相关联，以分析相关突变。基于这些突变，设计了一种利用高分辨率熔解（HRM）曲线分析的快速检测系统，并由深圳队列（由38株（）菌株组成）进行验证。基因组分析确定（）的2143等位基因从A突变为G（A2143G）是与CLR耐药性最相关的突变（P<0.01）。在HRM系统中，野生型（）的熔解温度（Tm）为79.28±0.01℃，而突变型的Tm为79.96±0.01℃。这些差异使得能够快速区分两种类型的（）（P<0.01）。验证检查表明，该系统能够在不受其他胃微生物影响的情况下，检测样品中低至0.005 ng/μL的目标DNA。该方法在深圳验证队列中也表现出良好的性能，准确率为81.58%，特异性为100%。我们开发了一种HRM系统，能够准确快速地检测（）中的CLR耐药性。该方法可直接用于胃微生物群样本的检测，为（）耐药性的简易检测提供了新的基准。
