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Multiplex Mediator Displacement Loop-Mediated Isothermal Amplification for Detection of Treponema pallidum and Haemophilus ducreyi.
Emerging Infectious Diseases ( IF 7.2 ) Pub Date : 2020-02-01 , DOI: 10.3201/eid2602.190505
Lisa Becherer , Sascha Knauf , Michael Marks , Simone Lueert , Sieghard Frischmann , Nadine Borst , Felix von Stetten , Sibauk Bieb , Yaw Adu-Sarkodie , Kingsley Asiedu , Oriol Mitjà , Mohammed Bakheit

Yaws, a neglected tropical disease caused by the bacterium Treponema pallidum subspecies pertenue, manifests as ulcerative skin lesions. Nucleic acid amplification tests, like loop-mediated isothermal amplification (LAMP), are versatile tools to distinguish yaws from infections that cause similar skin lesions, primarily Haemophilus ducreyi. We developed a novel molecular test to simultaneously detect T. pallidum and H. ducreyi based on mediator displacement LAMP. We validated the T. pallidum and H. ducreyi LAMP (TPHD-LAMP) by testing 293 clinical samples from patients with yaws-like lesions. Compared with quantitative PCR, the TPHD-LAMP demonstrated high sensitivity and specificity for T. pallidum (84.7% sensitivity, 95.7% specificity) and H. ducreyi (91.6% sensitivity, 84.8% specificity). This novel assay provided rapid molecular confirmation of T. pallidum and H. ducreyi DNA and might be suitable for use at the point of care. TPHD-LAMP could support yaws eradication by improving access to molecular diagnostic tests at the district hospital level.

中文翻译:

多重介质置换环介导的等温扩增检测梅毒螺旋体和杜克雷嗜血杆菌。

雅司病是一种由梅毒螺旋体亚种持续引起的被忽视的热带疾病,表现为溃疡性皮肤损伤。核酸扩增测试,如环介导等温扩增 (LAMP),是区分雅司病和引起类似皮肤损伤的感染(主要是杜克雷嗜血杆菌)的通用工具。我们开发了一种新的分子测试,可基于介质置换 LAMP 同时检测 T. pallidum 和 H. ducreyi。我们通过测试来自患有偏航样病变的患者的 293 个临床样本,验证了梅毒螺旋体和杜克雷伊灯 (TPHD-LAMP)。与定量 PCR 相比,TPHD-LAMP 对梅毒螺旋体(84.7% 的敏感性,95.7% 的特异性)和 H. ducreyi(91.6% 的敏感性,84.8% 的特异性)表现出较高的敏感性和特异性。这种新的检测方法提供了梅毒螺旋体和杜克雷伊螺旋体 DNA 的快速分子确认,可能适合在护理点使用。TPHD-LAMP 可以通过改善地区医院分子诊断测试的可及性来支持消除偏航。
更新日期:2020-01-21
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