Multiplex Mediator Displacement Loop-Mediated Isothermal Amplification for Detection of <i>Treponema pallidum</i> and <i>Haemophilus ducreyi</i>
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 A. Bakheit
Abstract
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.