Rahumatullah, Anizah
(2012)
Development Of Conventional Real-Time And Thermostabilised Multiplex PCR For The Detection Of Toxoplasma Gondii.
Masters thesis, Universiti Sains Malaysia.
Abstract
Toxoplasma gondii is an important pathogen in veterinary and human medicines. It is
widely distributed throughout the world and has a broad range of hosts that includes
warm-blooded animals as intermediate hosts and felid (cat) as the definitive host.
Infection with this parasite is usually asymptomatic in healthy individuals but can
cause morbidity and mortality in immunocompromised patients and in congenitally
infected infants. Molecular biology is increasingly being used as a diagnostic method
for diagnosis of toxoplasmosis, especially for specimens collected from amniotic fluid,
cerebrospinal fluid, placenta tissue, blood and eye fluid. However, the existing PCRbased
methods for the detection of Toxoplasma DNA suffer from variations in
performance among the laboratories. Indeed, the use of PCR-based assay for diagnosis
of Toxoplasma is still uncommon in Malaysia. The aim of this study is to develop
rapid, specific and sensitive PCR-based assays to detect T. gondii DNA for diagnosis
of toxoplasmosis. Three PCR-based assays namely conventional multiplex PCR, realtime
multiplex PCR and thermostabilised PCR for the detection of T. gondii DNA
were developed. All the primers and probes used were newly designed and optimized.
All three PCR assays showed 100% specificity whereby no cross-reactivity with other
organisms was observed.
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