Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Handheld Ultra-Fast duplex PCR assays and Lateral flow detection and identification of Leishmania parasites for cutaneous leishmanisis diagnosis

Insaf Bel Hadj Ali
* ORCID logo ,
Yusr Saadi Ben Aoun
,
Zeineb Hammami
,
Oumayma Rhouma
,
Ahmed Sahbi Chakroun
,
Ikram Guizani
Version 1 : Received: 2 October 2023 / Approved: 2 October 2023 / Online: 2 October 2023 (09:43:13 CEST)

A peer-reviewed article of this Preprint also exists.

Bel Hadj Ali, I.; Saadi-Ben Aoun, Y.; Hammami, Z.; Rhouma, O.; Chakroun, A.S.; Guizani, I. Handheld Ultra-Fast Duplex Polymerase Chain Reaction Assays and Lateral Flow Detection and Identification of Leishmania Parasites for Cutaneous Leishmaniases Diagnosis. Pathogens 2023, 12, 1292. Bel Hadj Ali, I.; Saadi-Ben Aoun, Y.; Hammami, Z.; Rhouma, O.; Chakroun, A.S.; Guizani, I. Handheld Ultra-Fast Duplex Polymerase Chain Reaction Assays and Lateral Flow Detection and Identification of Leishmania Parasites for Cutaneous Leishmaniases Diagnosis. Pathogens 2023, 12, 1292.

Abstract

: Early and accurate detection of infectious diseases is a key step for surveillance, epidemiology, and control, notably timely disease diagnosis, patient management and follow-up. In this study, we aimed to develop hand-held ultra-fast duplex PCR assays coupled to amplicon detection by lateral flow (LF) immunoassay to deliver a rapid, and simple molecular diagnostic test for con-comitant detection and identification of the main Leishmania parasites encountered in Tunisia and the Old World. We selected two DNA targets to amplify L. major/L. tropica and L. infantum/L. tropica group of species DNAs, respectively. We optimized the experimental conditions of a du-plex ultra-fast PCR. The amplification is performed by a portable Palm convection PCR machine within 18 min and the products are detected by a LF cassette within 10 minutes. The test allows the identification of the infecting species according to the position and number of test lines re-vealed. Tested on a selection of DNAs of representative Leishmania strains of the three studied species (N=37), the ultra-fast duplex PCR-LF showed consistent, stable, and reproducible results. The analytical limit of detection of the test was 0.4pg for L. major, 4pg for L. infantum and 40pg for L. tropica.

Keywords

Cutaneous leishmaniases; Molecular diagnosis; Point of care; Leishmania; Molecular target; Palm PCR; Duplex PCR; Lateral Flow immunoassay

Subject

Biology and Life Sciences, Biology and Biotechnology

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.

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