当前位置: X-MOL 学术Gut Pathog. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Evaluation of multiplex ARMS-PCR for detection of Helicobacter pylori mutations conferring resistance to clarithromycin and levofloxacin.
Gut Pathogens ( IF 4.3 ) Pub Date : 2020-07-10 , DOI: 10.1186/s13099-020-00373-6
Yangxia Li 1 , Tangshan Lv 1, 2 , Chaochao He 1, 2 , Hongyan Wang 1, 2 , David S Cram 1, 2 , Linfu Zhou 1, 3 , Jun Zhang 4 , Weiqin Jiang 5
Affiliation  

Helicobacter pylori bacterium is a major cause of gastritis. With increasing use of antibiotics to treat infections, mutation resistant strains have emerged in most human populations. To effectively treat patients to help resolve infections, the clinician needs information on the antibiotic susceptibility profile of the infection. Therefore, a rapid and accurate test is required to provide this information. To address this issue, we designed and validated a real time multiplex ARMS-PCR assay for rapid detection of highly prevalent H. pylori clarithromycin and levofloxacin resistance mutations. The aim of the study was to evaluate the analytical and diagnostic sensitivity and specificity of ARMS-PCR, using direct Sanger sequencing of the known resistance mutations as the gold standard. In preliminary studies using a defined number of plasmids with clarithromycin and levofloxacin resistance mutations, the analytical sensitivity of our ARMS-PCR assay was 50 plasmid copies, equating to around 50 bacterium in a gastric biopsy sample. In terms of specificity, the assay was highly specific for the targeted resistance mutations. The assay was also able to reliably and efficiently detect heteroresistance of clarithromycin and levofloxacin mutations, even at a disproportional ratio of 1:1000. From the analysis of 192 samples with suspected H. pylori infections, the diagnostic sensitivity and specificity of the assay was very high for detection of clarithromycin resistance (100% and 100%), levofloxacin resistance (98.04% and 95.04%) and clarithromycin and levofloxacin double resistance (100% and 96.91%). Amongst the 74 patients diagnosed antibiotic resistance bacteria, 23 (31.1%) had clarithromycin resistance, 21 (28.4%) had levofloxacin resistance and 30 (40.5%) had double resistance. From sample receipt to results, our single tube assay could be routinely completed in under 2 h. Our assay demonstrated high diagnostic sensitivity and specificity for detection of clarithromycin and levofloxacin resistant H. pylori. Based on proven accuracy, together with high efficiency, scalability and low cost, our assay has useful clinical utility for rapid diagnosis of clarithromycin and levofloxacin resistant H. pylori infections. Our assay results will provide patients with a clear diagnosis, enabling the treating clinician to administer the most effective antibiotic regimen to help the clear the infection.

中文翻译:

评估多重 ARMS-PCR 检测幽门螺杆菌突变导致对克拉霉素和左氧氟沙星的耐药性。

幽门螺杆菌是胃炎的主要原因。随着越来越多地使用抗生素治疗感染,大多数人群中出现了抗突变菌株。为了有效地治疗患者以帮助解决感染,临床医生需要有关感染的抗生素敏感性特征的信息。因此,需要快速准确的测试来提供此信息。为了解决这个问题,我们设计并验证了一种实时多重 ARMS-PCR 检测方法,用于快速检测高度流行的幽门螺杆菌克拉霉素和左氧氟沙星耐药突变。该研究的目的是评估 ARMS-PCR 的分析和诊断敏感性和特异性,使用已知抗性突变的直接 Sanger 测序作为金标准。在使用确定数量的具有克拉霉素和左氧氟沙星耐药突变的质粒的初步研究中,我们的 ARMS-PCR 测定的分析灵敏度为 50 个质粒拷贝,相当于胃活检样本中约 50 个细菌。在特异性方面,该测定对靶向抗性突变具有高度特异性。该测定还能够可靠有效地检测克拉霉素和左氧氟沙星突变的异抗性,即使在不成比例的比例为 1:1000 时也是如此。通过对 192 份疑似幽门螺杆菌感染的样本的分析,该方法对克拉霉素耐药(100% 和 100%)、左氧氟沙星耐药(98.04% 和 95.04%)以及克拉霉素和左氧氟沙星的检测具有很高的诊断敏感性和特异性双电阻(100% 和 96.91%)。74例确诊为耐药菌的患者中,克拉霉素耐药23例(31.1%),左氧氟沙星耐药21例(28.4%),双重耐药30例(40.5%)。从样品接收到结果,我们的单管检测通常可以在 2 小时内完成。我们的测定显示了对克拉霉素和左氧氟沙星耐药幽门螺杆菌的高诊断敏感性和特异性。基于经过验证的准确性,以及高效率、可扩展性和低成本,我们的检测在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。1%)对克拉霉素耐药,21名(28.4%)对左氧氟沙星耐药,30名(40.5%)对双重耐药。从样品接收到结果,我们的单管检测通常可以在 2 小时内完成。我们的测定显示了对克拉霉素和左氧氟沙星耐药幽门螺杆菌的高诊断敏感性和特异性。基于经过验证的准确性,以及高效率、可扩展性和低成本,我们的检测在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。1%)对克拉霉素耐药,21名(28.4%)对左氧氟沙星耐药,30名(40.5%)对双重耐药。从样品接收到结果,我们的单管检测通常可以在 2 小时内完成。我们的测定显示了对克拉霉素和左氧氟沙星耐药幽门螺杆菌的高诊断敏感性和特异性。基于经过验证的准确性,以及高效率、可扩展性和低成本,我们的检测在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。从样品接收到结果,我们的单管检测通常可以在 2 小时内完成。我们的测定显示了对克拉霉素和左氧氟沙星耐药幽门螺杆菌的高诊断敏感性和特异性。基于经过验证的准确性,以及高效率、可扩展性和低成本,我们的检测在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。从样品接收到结果,我们的单管检测通常可以在 2 小时内完成。我们的测定显示了对克拉霉素和左氧氟沙星耐药幽门螺杆菌的高诊断敏感性和特异性。基于经过验证的准确性,以及高效率、可扩展性和低成本,我们的检测在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。结合高效率、可扩展性和低成本,我们的检测方法在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。结合高效率、可扩展性和低成本,我们的检测方法在快速诊断克拉霉素和左氧氟沙星耐药性幽门螺杆菌感染方面具有有用的临床实用性。我们的检测结果将为患者提供明确的诊断,使治疗的临床医生能够使用最有效的抗生素治疗方案来帮助清除感染。
更新日期:2020-07-10
down
wechat
bug