Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in yaws-like lesions

Michael Frimpong
* ORCID logo ,
Shirley Victoria Simpson
,
Hubert Senanu Ahor
,
Abigail Agbanyo
,
Solomon Gyabaah
ORCID logo ,
Bernadette Agbavor
ORCID logo ,
Ivy Brago Amanor
,
Kennedy Kwasi Addo
,
Susanne Böhlken-Fascher
,
Jonas Kissenkötter
,
Ahmed Abd El Wahed
ORCID logo ,
Richard Odame Phillips
Version 1 : Received: 25 August 2020 / Approved: 26 August 2020 / Online: 26 August 2020 (09:01:07 CEST)

A peer-reviewed article of this Preprint also exists.

Frimpong, M.; Simpson, S.V.; Ahor, H.S.; Agbanyo, A.; Gyabaah, S.; Agbavor, B.; Amanor, I.B.; Addo, K.K.; Böhlken-Fascher, S.; Kissenkötter, J.; Wahed, A.A.E.; Phillips, R.O. Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions. Trop. Med. Infect. Dis. 2020, 5, 157. Frimpong, M.; Simpson, S.V.; Ahor, H.S.; Agbanyo, A.; Gyabaah, S.; Agbavor, B.; Amanor, I.B.; Addo, K.K.; Böhlken-Fascher, S.; Kissenkötter, J.; Wahed, A.A.E.; Phillips, R.O. Multiplex Recombinase Polymerase Amplification Assay for Simultaneous Detection of Treponema pallidum and Haemophilus ducreyi in Yaws-Like Lesions. Trop. Med. Infect. Dis. 2020, 5, 157.

Abstract

Yaws is a skin debilitating disease caused by Treponema pallidum subspecies pertenue with most cases reported in children. World Health Organization (WHO) aims at total eradication of this disease through mass treatment of suspected cases followed by an intensive follow-up program. However, effective diagnosis is pivotal in the successful implementation of this control program. Recombinase polymerase amplification (RPA), an isothermal nucleic acid amplification technique offers a wider range of differentiation of pathogens including those isolated from chronic skin ulcers with similar characteristics such as Haemophilus ducreyi (H. ducreyi). We have developed a duplex RPA assay for the simultaneous detection of Treponema pallidum (T. pallidum) and H. ducreyi (TPHD-RPA). TPHD-RPA assay demonstrated no cross-reaction with other pathogens and enable detection of T. pallidum and H. ducreyi within 15 minutes at 42 oC. The duplex RPA assay was validated with 49 clinical samples from individuals confirmed to have yaws by serological tests. Compared with commercial multiplex real-time PCR, the TPHD-RPA assay demonstrated 94-95% sensitivity for T. pallidum and H. ducreyi confirmed samples, respectively and 100% specificity. This simple novel TPHD-RPA assay enables the rapid detection of both T. pallidum and H. ducreyi in yaws-like lesions. This test could support the yaws eradication programs by ensuring effective diagnosis as well as enable monitoring of eradication efforts success or failure and planning of follow-up interventions at the community level.

Keywords

Recombinase polymerase amplification; Treponema pallidum; Haemophilus ducreyi; Molecular diagnostics; Point-of-care test

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Download PDF

Comments (0)

We encourage comments and feedback from a broad range of readers. See criteria for comments and our Diversity statement.

Leave a public comment
Send a private comment to the author(s)
* All users must log in before leaving a comment
Views 0
Downloads 0
Comments 0
Metrics 0
Get PDF Cite Share 0
Bookmark BibSonomy Mendeley Reddit Delicious
×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here.