|
Women in Science Mentoring Program (2018)
SG-007: Point-of-care cutaneous leishmaniasis diagnosis: handheld fast PCR assays and lateral flow detection for Leishmania parasites detection and identification
PI: Insaf Bel Hadj Ali, Institut Pasteur de Tunis
Project Dates: September 2019 - August 2021
Project Overview
Early and accurate detection of infectious diseases is a key step not only for surveillance, epidemiology, and control but also, notably, for timely disease diagnosis, patient management, and follow-up. In recent decades, several diagnostic tests satisfying these criteria were developed to identify major human pathogens such as human immunodeficiency virus (HIV), tuberculosis (TB), and malaria, as these diseases received much more attention compared to neglected tropical diseases such as leishmaniases. Leishmaniases are a group of vector-born parasitic diseases with a wide range of clinical manifestations, some of which are fatal if left untreated. More than one million cutaneous leishmaniases (CL) cases are annually reported, with 80% of those occurring in the Middle East and North Africa (MENA) region. In addition, the primary drugs employed for CL treatment are toxic, and their efficiency may depend on parasite species/strains. CL diagnosis is also complicated, as it is routinely performed via microscopy direct examination, a time-consuming technique that requires trained personnel and cannot identify particular parasites.
This PEER project aimed to identify novel CL molecular diagnosis assays that satisfy criteria for timely patient management and disease control and equip areas with low resources and poor laboratory infrastructure with equitable access to high-quality patient diagnosis and management. This project also supported the scientific training and mentoring of several female researchers under the guidance of the PI Dr. Ali, leading to completed research and degrees, as well as enhanced future research goals.
Final Summary of Project Activities
This PEER Women’s Mentorship seed grant arose out of the PI Dr. Ali’s work as a part of PEER project 5-518, during which she and her colleagues identified five promising targets to use in fast PCR assays for leishmaniases. All targets included sequence information covering different strains of the species of interest. Within these targets, they designed four primer pairs targeting the L. major /L. Tropica group of species and two targeting the L. infantum /L. tropica group of species. Five markers gave specific profiles (mt22F1R1, mt22F2R2, mt30F/R, it20F1/R1, and it20F2/R2) as expected and were therefore selected for the fast and ultrafast PCR assays development. The researchers set up and processed several types of PCR assays with different types and processing times, including simplex fast PCR, fast duplex PCR, simplex ultra-fast PCR, and duplex ultra-fast PCR. They tested the detection limit of various Leishmania strains and the impact of human DNA on the assays. Among their findings was the ultra-fast duplex PCR showed consistent, stable, and reproducible results.
This follow-on seed grant project supported the continuing research and mentoring of several female scientists, including a doctoral student, a biology student, and an engineering student. As part of the research team, they worked with the PI on strategy and methodology of the study and received training on molecular biology techniques, as well as bioinformatic analyses. The PI also established a mentoring program where she met individually and in monthly group meetings with young scientists. The meetings included discussions of their short- and long-term scientific and educational goals, as well as identifying goals in technical skills development, leadership skills improvement, personal branding, work-life balance, and career development.
The PEER team also held a workshop on technologies based on Palm PCR and lateral flow PCR attended by parasitologists, lab technicians, and students. The PI also received an additional grant to continue her work.
Publication
Insaf Bel Hadj Ali, Yusr Saadi-Ben Aoun, Zeineb Hammami, Oumayma Rhouma, Ahmed Sahbi Chakroun, and Ikram Guizani. 2023. Handheld ultra-fast duplex polymerase chain reaction assays and lateral flow detection and identification of Leishmania parasites for cutaneous leishmaniases diagnosis. Pathogens 12(11): 1292. https://doi.org/10.3390/pathogens12111292
Back to WSMP Grant Recipients
|
|
|
|
