Guan Ai-Xing, Yang Shuang-Yan, Wu Tong, Zhou Wen-Ting, Chen Hao, Huang Zan-Song, Luo Pei-Pei, Huang Yan-Qiang
Department of Gastroenterology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, Guangxi Zhuang Autonomous Region, China.
Guangxi Clinical Medical Research Center for Hepatobiliary Diseases, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, Guangxi Zhuang Autonomous Region, China.
World J Gastroenterol. 2025 Aug 28;31(32):106424. doi: 10.3748/wjg.v31.i32.106424.
(), a globally prevalent pathogen, is exhibiting increasing rates of antimicrobial resistance. However, clinical implementation of pre-treatment susceptibility testing remains limited due to the organism's fastidious growth requirements and prolonged culture time.
To propose a novel detection method utilizing antibiotic-supplemented media to inhibit susceptible strains, while resistant isolates were identified through urease-mediated hydrolysis of urea, inducing a phenol red color change for visual confirmation.
Colombia agar was supplemented with urea, phenol red, and nickel chloride, and the final pH was adjusted to 7.35. Antibiotic-selective media were prepared by incorporating amoxicillin (0.5 μg/mL), clarithromycin (2 μg/mL), metronidazole (8 μg/mL), or levofloxacin (2 μg/mL) into separate batches. Gastric antral biopsies were homogenized and inoculated at 1.0 × 10 CFU onto the media, and then incubated under microaerobic conditions at 37 °C for 28-36 hours. Resistance was determined based on a color change from yellow to pink, and the results were validated broth microdilution according to Clinical and Laboratory Standards Institute guidelines.
After 28-36 hours of incubation, the drug-resistant isolates induced a light red color change in the medium. Conversely, susceptible strains ( 26695 and G27) produced no visible color change. Compared with the conventional 11-day protocol, the novel method significantly reduced detection time. Among 201 clinical isolates, 182 were successfully evaluated using the new method, resulting in a 90.5% detection rate. This was consistent with the 95.5% agreement rate observed when compared with microdilution-based susceptibility testing. The success rate of the novel approach was significantly higher than that of the comparative method ( < 0.01). The accuracy of the new method was comparable to that of the dilution method.
The novel detection method can rapidly detect drug resistance within 28-36 hours. With its operational simplicity and high diagnostic performance, it holds strong potential for clinical application in the management of antimicrobial resistance.
（某种病原体）是一种全球流行的病原体，其耐药率正在上升。然而，由于该生物体苛求的生长要求和较长的培养时间，治疗前药敏试验的临床应用仍然有限。
提出一种新的检测方法，利用添加抗生素的培养基抑制敏感菌株，而耐药菌株则通过脲酶介导的尿素水解来鉴定，尿素水解会导致酚红颜色变化以便目视确认。
在哥伦比亚琼脂中添加尿素、酚红和氯化镍，并将最终pH值调至7.35。通过将阿莫西林（0.5μg/mL）、克拉霉素（2μg/mL）、甲硝唑（8μg/mL）或左氧氟沙星（2μg/mL）分别加入不同批次来制备抗生素选择性培养基。将胃窦活检组织匀浆并以1.0×10CFU接种到培养基上，然后在37℃微需氧条件下孵育28 - 36小时。根据颜色从黄色变为粉红色来确定耐药性，结果根据临床和实验室标准协会指南通过肉汤微量稀释法进行验证。
孵育28 - 36小时后，耐药菌株在培养基中引起浅红色颜色变化。相反，敏感菌株（26695和G27）没有产生可见的颜色变化。与传统的11天方案相比，新方法显著缩短了检测时间。在201株临床分离株中，182株使用新方法成功评估，检测率为90.5%。这与基于微量稀释的药敏试验相比观察到的95.5%的符合率一致。新方法的成功率显著高于比较方法（<0.01）。新方法的准确性与稀释法相当。
这种新的检测方法可以在28 - 36小时内快速检测耐药性。因其操作简单和高诊断性能，在抗菌药物耐药性管理的临床应用中具有很强的潜力。
