Multiplex Mediator Displacement Loop-Mediated Isothermal Amplification for Detection of Treponema pallidum and Haemophilus ducreyi

Emerg Infect Dis. 2020 Feb;26(2):282-288. doi: 10.3201/eid2602.190505.

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.

Keywords: Haemophilus ducreyi; LAMP; PCR; Treponema pallidum; Yaws; bacteria; loop-mediated isothermal amplification; neglected tropical diseases.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chancroid / diagnosis*
  • Chancroid / microbiology
  • Child
  • Female
  • Ghana
  • Haemophilus ducreyi / isolation & purification*
  • Humans
  • Male
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques
  • Papua New Guinea
  • Sensitivity and Specificity
  • Treponema pallidum / isolation & purification*
  • Yaws / diagnosis*
  • Yaws / microbiology

Supplementary concepts

  • LAMP assay