Basuni, Madihah
(2012)
Multiplex Real-Time PCR For The Detection Of Pathogenic Intestinal Parasites And Comparison With Parasitological Techniques.
Masters thesis, Universiti Sains Malaysia.
Abstract
Intestinal parasitic infections by helminths and protozoa are among the most
prevalent infections and remain a major public health burden in underdeveloped
countries. Most intestinal helminth infections cause abdominal pain accompanied by
aneroxia, nausea and diarrhea while intestinal protozoa cause diarrheal diseases.
Routine diagnostic methods for intestinal parasitic infections which rely heavily on
microscopic detection are insensitive, and require well-trained microscopists to avoid
misdiagnosis or overdiagnosis of the infection. These limitations have led to the
development of highly sensitive DNA-based assays. PCR has been proven to be
sensitive and specific for detection of enteric pathogens. Since conventional PCR is
time consuming and prone to cross-contamination, it is desirable to develop a realtime
PCR assay which is rapid and can provide quantitative and real-time
information on the amplified products.
In this study, a real-time multiplex PCR assay for the detection of closely related
intestinal helminths namely Ascaris lumbricoides, Strongyloides stercoralis, Necator
americanus and Ancylostoma duodenale and an assay for the detection of three
closely related intestinal protozoa which comprised Entamoeba histolytica,
Cryptosporidium parvum and Giardia lamblia were optimized and evaluated.
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