molecular detection and identification of parasites involved in human disease
abstract
the detection and identification of disease-causing parasites via dna and the
polymerase chain reaction (pcr) offers a new approach for quick and efficient disease diagnosis.
three nucleic acid based technologies have been developed and optimized for the identification
of six species of parasites that cause disease, and designed to amplify small fragments of dna
likely to remain in archaeological material. two methods involve a species-specific
identification by examining four gene fragments, one of which has been designed as a multiplex
pcr to minimize the time required for diagnosis. the third method involves the use of one
species-specific gene target per parasite in a multiplex pcr for the detection of multiple
parasites in one reaction to expedite the testing process and confirmation of the disease in
question in a clinical setting. sensitivity and specificity have been proven through the use of a
1/1000 diluted dna template and sequencing of products confirmed the presence o f each
species analyzed. with increased sensitivity, a less invasive sampling procedure would be
required from the patient. also, the earlier the presence of parasites in the tissues is detected, the
earlier the diagnosis would occur. tissues that contain very few parasites and had been
classified as previously unusable for diagnosis or tissues from older medical cases can be
analyzed for the presence of disease and the identification of causative species. the successful
application of the singleplex pcr technique to plasmodium falciparum and leishmania
degraded dna samples leads to the feasibility of success for the multiplex pcr techniques
involving degraded dna analyses. therefore, this methodology may be applied to samples of
unknown disease content to either include or exclude the agents of disease. future directions
may extend these analyses to include a range of other agents of disease, thus allowing for the
determination of disease antiquity through degraded dna analyses.
collections
- retrospective theses [1604]