Use of molecular and bioinformatics tools for developing methods of epidemiological identification of trypanosomes
type: CreativeWork
description: Human African trypanosomiasis (HAT), also known as sleeping sickness, has been a major health problem for populations in Africa caused by the Trypanosoma brucei spp parasite. Although, the current number of the reported cases is on the decrease, more efforts are required to try to control or eliminate the disease. The recent advances in molecular techniques have contributed towards identifying taxonomic groups at all levels (species, subspecies, populations, strains and isolates). Commonly, field samples are collected and stored using Whatman FTA cards. Many molecular epidemiological tools are available for detection and strain typing in trypanosomes. These tools include nested ITS-PCR, which is based on size variation of the ITS genes and MGE-PCR, which is based on variations in position of mobile genetic elements (MGEs). Although commonly used, these tools have not been fully validated. For example, the ITS-PCR has not been used or validated against blood samples obtained from sleeping sickness patients in Angola. Furthermore, the MGE-PCR system has not been evaluated for use directly from FTA cards.The aim of this thesis is to develop improved molecular tools to assist diagnostic and epidemiological studies. In order to improve the molecular diagnostic use of Whatman FTA cards, an extraction method based on Chelex was investigated. Using Chelex to extractT. brucei DNA from FTA cards, followed by a nested ITS-PCR detection system, allowed parasite DNA detection to 1ng/µl. To evaluate this tool on field samples, ITS-PCR amplification was carried on DNA eluted by Chelex extraction from 36 FTA cards spotted with blood from Angolan patients tested positive for trypanosomiasis by the card agglutination test (CATT). Twenty four of these samples were successfully PCR amplified using mammalian tubulin primers. Of these 24 samples, 11 (= 45.8%) were confirmed as trypanosome positive utilising a specific ITS-PCR based approach. As such, this indicates that further work is n…
abstract: Human African trypanosomiasis (HAT), also known as sleeping sickness, has been a major health problem for populations in Africa caused by the Trypanosoma brucei spp parasite. Although, the current number of the reported cases is on the decrease, more efforts are required to try to control or eliminate the disease. The recent advances in molecular techniques have contributed towards identifying taxonomic groups at all levels (species, subspecies, populations, strains and isolates). Commonly, field samples are collected and stored using Whatman FTA cards. Many molecular epidemiological tools are available for detection and strain typing in trypanosomes. These tools include nested ITS-PCR, which is based on size variation of the ITS genes and MGE-PCR, which is based on variations in position of mobile genetic elements (MGEs). Although commonly used, these tools have not been fully validated. For example, the ITS-PCR has not been used or validated against blood samples obtained from sleeping sickness patients in Angola. Furthermore, the MGE-PCR system has not been evaluated for use directly from FTA cards.The aim of this thesis is to develop improved molecular tools to assist diagnostic and epidemiological studies. In order to improve the molecular diagnostic use of Whatman FTA cards, an extraction method based on Chelex was investigated. Using Chelex to extractT. brucei DNA from FTA cards, followed by a nested ITS-PCR detection system, allowed parasite DNA detection to 1ng/µl. To evaluate this tool on field samples, ITS-PCR amplification was carried on DNA eluted by Chelex extraction from 36 FTA cards spotted with blood from Angolan patients tested positive for trypanosomiasis by the card agglutination test (CATT). Twenty four of these samples were successfully PCR amplified using mammalian tubulin primers. Of these 24 samples, 11 (= 45.8%) were confirmed as trypanosome positive utilising a specific ITS-PCR based approach. As such, this indicates that further work is n…
author: Adwik, GA
language: en
date published: 2026
date modified: 2026-07-24T04:26:44Z
provider: U. of Salford
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: U. of Salford