A Polymerase chain reaction (PCR) based assay for the detection of DIROFILARIA REPENS

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dc.contributor.authorABAYASEKARA, SNWen_US
dc.date.accessioned2011-02-14T15:13:20Z
dc.date.available2011-02-14T15:13:20Z
dc.date.created1997en_US
dc.date.issued1997en_US
dc.descriptionDissertation: M.Sc., University of Colombo: UC(MED), 1997.en_US
dc.description.abstractFilariasis is a group of infectious diseases caused by parasitic nematodes. Of the many species of filarial worms eight are truly specific for man, but there are also zoonotic forms which can infect humans. Dirofilaria repens is one such parasitic nematode of dogs in Sri Lanka. According to current estimates, 410 cases of human dirofilariasis caused by Dirofilaria repens has been reported exclusively in the old world including Sri Lanka. Given the low sensitivity of current diagnostic methods (blood smears) and carying expertise between laboratories and technicians, a rapid and sensitive assay would greatly assist the early detection of microfilariae in dogs and developing infective larvae in vectors. The capability to detect genetic elements (DNA or RNA) of a particular parasite or organism as a means of identifying the causative agent has been the traditional function of nucleic acid hybridization assays. However the low copy number of genetic material and the need for using radiolabelling fostered the development of PCR based assays. A previously cloned and characterized tandemly repeated DNA sequence of D.repens was selected for designing primers for the development of a PCR based assay. The primer pair (DRF4 \& DRR2) of three pairs of primers was chosen for the assay. The PCR assay was optimized in relation to the concentration of deoxyribonucleotides (dNTP), MgCl2, primers, Taq polymerase and annealing temperature. The assay selectively amplified D.repens DNA only and was sensitive enough to detect 100 fg of parasite DNA and a single microfilaria in infected blood samples. Analysis of blood samples from dogs (n=50) using the PCR assay was compared with that of microscopy. The results obtained by PCR were in total agreement with that of microscopy, except for one sample which was negative. It is likely that this sample may have had a very low level of microfilariaemia and hence not detected by microscopy. The high sensitivity of the assay could perhaps also aid in the identification of immature adults found as lumps or nodules in human clinical specimens obtained from biopsiesen_US
dc.identifier.citationABAYASEKARA, SNW, A Polymerase chain reaction (PCR) based assay for the detection of DIROFILARIA REPENS, University of Colombo UC(MED), 1997: 103p.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/129459
dc.language.isoen_USen_US
dc.publisherUniversity of Colombo: UC(MED).en_US
dc.rightsUniversity of Colombo, UC(MED): Sri Lanka HELLIS Networken_US
dc.source.urihttps://hellis.srilanka.healthrepository.orgen_US
dc.subjectFilarioideaen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshDirofilariaen_US
dc.subject.meshDirofilariasisen_US
dc.subject.meshDirofilaria-isolation \& purificationen_US
dc.subject.meshDirofilariasis-geneticsen_US
dc.subject.meshDirofilariasis-diagnosisen_US
dc.subject.meshDirofilariasis-parasitologyen_US
dc.titleA Polymerase chain reaction (PCR) based assay for the detection of DIROFILARIA REPENSen_US
dc.typeThesisen_US
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