Ren Xinlu, Suo Baojun, Li Cailing, Ping Guangjie, Ma Lingling, Shi Yanyan, Zhou Kai, Wang Yuxin, Tian Xueli, Zhou Liya, Song Zhiqiang
Department of Gastroenterology, Peking University Third Hospital, Beijing, China.
Research Center of Clinical Epidemiology, Peking University Third Hospital, Beijing, China.
J Clin Microbiol. 2025 Jan 31;63(1):e0103424. doi: 10.1128/jcm.01034-24. Epub 2024 Dec 16.
Genotypic methods for detecting antibiotic resistance in infection offer a rapid, convenient, and accurate approach for tailored therapy. However, existing studies predominantly examine single sample types and lack comparative analyses across different samples. This study comprehensively detects and compares genotypic resistance to clarithromycin and levofloxacin in gastric mucosa, gastric fluid, and fecal samples from the same patients. The study enrolled 183 participants, comprising 124 . -positive and 59 . -negative patients. All participants provided fecal samples and underwent gastroscopy for the collection of gastric mucosa and gastric fluid. Real-time PCR was employed to detect genotypic resistance to clarithromycin and levofloxacin in conjunction with bacterial culture and antibiotic susceptibility testing. Genotypic resistance detection rates for clarithromycin were 100% in gastric mucosa, 99.2% in gastric fluid, and 79.8% in fecal samples. For levofloxacin, detection rates were 97.6%, 96.8%, and 72.6%, respectively. The results showed that PCR detection for clarithromycin exhibited high sensitivity (0.94-0.95) and specificity (0.88-0.89) across all sample types. However, PCR detection for levofloxacin demonstrated slightly lower sensitivity (0.79-0.89) and specificity (0.79-0.83). The comparison of genotypic resistance results by PCR among the three sample types showed that gastric mucosa and gastric juice exhibited higher consistency, while the consistency between feces and both gastric mucosa and gastric juice was lower. This study confirmed good consistency between genotypic and phenotypic resistance in clarithromycin and levofloxacin. While both gastric mucosa and gastric fluid samples demonstrated high detection performance, the efficiency of detecting fecal samples was constrained by challenges in DNA extraction.
This study, with a large sample size, comprehensively tested both pylori-negative and -positive patients, including rapid urease test, histopathological evaluation and staining, bacterial culture, susceptibility testing, and resistance gene mutation analysis. By simultaneously examining gastric mucosa, gastric juice, and fecal samples from the same individuals, we minimized confounding factors arising from different sample sources, ensuring the reliability of our results. This approach effectively delineated the differences and characteristics in detection performance among different sample types, offering crucial reference data for selecting appropriate detection samples and identifying areas for improvement. The findings revealed robust concordance between genotypic and phenotypic resistance, with both gastric mucosa and gastric juice samples demonstrating excellent detection performance. However, the efficiency of detecting resistance in fecal samples was hampered by challenges in DNA extraction.
用于检测感染中抗生素耐药性的基因分型方法为个性化治疗提供了一种快速、便捷且准确的途径。然而,现有研究主要检测单一样本类型,缺乏对不同样本的比较分析。本研究全面检测并比较了同一患者的胃黏膜、胃液和粪便样本中对克拉霉素和左氧氟沙星的基因分型耐药性。该研究纳入了183名参与者,包括124名幽门螺杆菌阳性和59名幽门螺杆菌阴性患者。所有参与者均提供粪便样本,并接受胃镜检查以收集胃黏膜和胃液。采用实时聚合酶链反应(PCR)结合细菌培养和抗生素敏感性试验来检测对克拉霉素和左氧氟沙星的基因分型耐药性。克拉霉素的基因分型耐药性检测率在胃黏膜中为100%,在胃液中为99.2%,在粪便样本中为79.8%。对于左氧氟沙星,检测率分别为97.6%、96.8%和72.6%。结果表明,针对克拉霉素的PCR检测在所有样本类型中均表现出高敏感性(0.94 - 0.95)和特异性(0.88 - 0.89)。然而,针对左氧氟沙星的PCR检测敏感性(0.79 - 0.89)和特异性(0.79 - 0.83)略低。三种样本类型之间通过PCR进行的基因分型耐药性结果比较显示,胃黏膜和胃液的一致性较高,而粪便与胃黏膜和胃液之间的一致性较低。本研究证实了克拉霉素和左氧氟沙星基因分型耐药性与表型耐药性之间具有良好的一致性。虽然胃黏膜和胃液样本均表现出高检测性能,但粪便样本的检测效率受到DNA提取挑战的限制。
本研究样本量较大,全面检测了幽门螺杆菌阴性和阳性患者,并进行了快速尿素酶试验、组织病理学评估和染色、细菌培养、药敏试验以及耐药基因突变分析。通过同时检测同一个体的胃黏膜、胃液和粪便样本,我们最大限度地减少了不同样本来源产生的混杂因素,确保了结果的可靠性。这种方法有效地描绘了不同样本类型在检测性能方面的差异和特点,为选择合适的检测样本和确定改进领域提供了关键参考数据。研究结果显示基因分型耐药性与表型耐药性之间具有很强的一致性,胃黏膜和胃液样本均表现出出色的检测性能。然而,粪便样本中耐药性检测的效率受到DNA提取挑战的阻碍。