Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item Polymerase Chain Reaction and mosquito dissection as tools to monitor filarial Infection levels following mass treatment in Gampaha District, Sri Lanka(Elsevier, 2008) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Bandara, K.B.A.T.; Liyanage, T.; Abeyewickreme, W.BACKGROUND: Mass Drug Administration (MDA)-based Global Lymphatic filariasis (Lf) eradication programmes are aimed at stopping transmission of Wuchereria bancrofti by its mosquito vector. The study was designed to compare one year post treatment (mass distribution of Diethylcarbamazine-Albendazole) infection rates of Wuchereria bancrofti in Culex quenquifaciatus, the main vector of Lf in Sri Lanka using Conventional dissection techniques and a Polymerase Chain Reaction (PCR) assay based on parasite specific Ssp1 repeat which amplifies a fragment of 188 bp. METHODS: Field study was conducted in 45 sites in all Medical Officer of Health (MOH) areas in the Gampaha district, Sri Lanka; identified by the Anti Filariasis Campaign (AFC) as having high-risk for bancroftian filariasis transmission. Indoor-resting mosquitoes were collected by aspiration from 20 houses per each site. Part of the mosquitos were used for dissection and the remainder was used for PCR to detect the filarial parasites in mosquito. RESULTS: Mosquito dissection data revealed 42.22% (19/45) of the sites were infested with mosquitoes positive for Wuchereria bancrofti, indicating 8 transmission active MOH areas (53.33%; 8/15). An infection rate of 5.26% was observed among the mosquitoes caught from households and the larval density was 8.7 per positive mosquito. PCR investigation revealed that 46.67% (21/45) of the sites were positive for W. bancrofti DNA, indicating 11 transmission active areas (73.33%; 11/15). The PCR was found to be more sensitive compared to microscopy in detecting the filarial parasite in field collected mosquito samples with respect to the MOH areas. CONCLUSION: The PCR technique employed offers scope for detection of the filarial parasites with higher sensitivity and specificity; is efficient and rapid. This technique applied for the first time in Sri Lanka, can be adopted as a diagnostic tool for the detection of filarial parasites in the vector population in surveillance to enable effective control of filariasis in the country. Acknowledgements: Authors acknowledge the WHO/SEARO/TDR (grant no. SN 1152) and University of Kelaniya (Research grant no. RP/03/04/06/01/2006) and to Ms. H.M.Renuka and Mr. H.P.Anura U. Pathirana, Mr. M.I.M.Peris and Mr. Y.L.Rassapana for their support during field study activities. © 2008 Elsevier Inc.Item A Comparative retrospective study of novel Reverse-Transcription Polymerase Chain Reaction-based Liquid Hybridization (RT-PCR-LH) assay with Polymerase Chain Reaction (PCR) amplification, virus isolation and serological techniques for early, definitive laboratory diagnosis of dengue infection(Malaysian Society of Parasitology and Tropical Medicine, 2007) Hapugoda, M.D.; de Silva, N.R.; Khan, B.; Gunasena, S.; Dayanath, M.Y.D.; Abeyewickreme, W.Dengue is an important vector borne viral infection in South East Asia. Dengue virus is responsible for dengue fever, dengue haemorrhagic fever and dengue shock syndrome. Early diagnosis of infection helps in monitoring the disease, determining when hospital admission is necessary and in reducing case fatalities. The objective of the study was to carry out a comparative retrospective study of a novel Reverse Transcription-Polymerase Chain Reaction-based Liquid Hybridization (RT-PCR-LH) assay with PCR amplification, virus isolation and serological techniques for laboratory diagnosis of dengue infection. Amplified products of Non Structural-3 gene were hybridized with a mixture of the 4 dengue type-specific Deoxyribonucleic Acid (DNA) probes in liquid phase. The assay was validated in a comparative retrospective study using acute serum samples collected from 88 patients with dengue confirmed by Haemagglutination Inhibition (HAI) assay. The assay was highly specific for diagnosis of dengue infection. As an early (<5 days of fever) laboratory diagnostic method, this assay had 100% sensitivity for detection of dengue patients confirmed by HAI assay. A high analytical sensitivity of 2 fluorescent focus units of dengue virus/reaction was achieved. Novel RT-PCR-LH assay using a single serum specimen offers distinct advantages of specificity and sensitivity over other diagnostic techniques for early definitive laboratory diagnosis of dengue infection at the time during which serological methods cannot be used.Item Confirmation of 2006 chikungunya outbreak in Sri Lanka using RT-PCR(Malaysian Society of Parasitology and Tropical Medicine, 2007) Abeyewickreme, W.; Bandara, K.B.A.T.; Perera, H.; Dayanath, M.Y.D.; Hapuarachchi, C.Chikungunya, a mosquito-borne viral infection caused by a single-stranded RNA virus of the family Togaviridae, is considered as a rare, non-fatal disease. During February to October 2006, an epidemic of over 1.3 million suspected cases was reported in India and neighbouring countries causing a significant economic loss due to crippling manifestations of this infection. With the outbreak of many viral fevers including dengue and dengue haemorrhagic fever, in October–November 2006, patients with manifestations suggestive of chikungunya such as high fever, headache, arthralgia and arthiritis (particularly, in ankle, knee and small joints of hands) were reported in many parts of Sri Lanka. As no chikungunya cases had been officially reported in the island since 1969, laboratory investigations for the presence of chikungunya virus was a prime requirement for confirmation of the outbreak. A total of 60 venous blood samples collected from suspected patients from different geographical regions of Sri Lanka were analysed using a reverse transcriptase-polymerase chain reaction (RT-PCR) technique to confirm the presence of chikungunya virus. Viral RNA was extracted from samples collected within 1-4 days of fever by using a Qiagen RNA extraction kit. RT-PCR was performed using chikungunya specific oligonucleotides. Both positive and negative controls were included in each set of reactions. The amplified products (354 bp) were visualized by running in a 1.5% agarose gel followed by ethidium bromide staining. Of the 60 samples, 33 (55%) were positive for chikungunya. They were distributed among almost all the geographical regions, highlighting the presence of a wide-spread epidemic in the country.Item A Comparative field study of novel commercial Antigen Detection Enzyme-Linked Immunosorbent Assay (ELISA) with Reverse Transcription Polymerase Chain Reaction (RT- PCR) assay for early definitive laboratory diagnosis of dengue viral infection in Sri Lanka(Sri Lanka Association for the Advancement of Science, 2007) Hapugoda, M.D.; Jayasooriya, D.H.S.W.; Gunawardene, Y.I.N.S.; Wellawaththage, C.; Premaratna, R.; Abeyewickreme, W.Dengue is an important mosquito borne viral infection in South East Asia. Early definitive laboratory diagnosis of infection would help in management of patients and reducing the case fatality rate. The objective of this study was to determine the accuracy of novel commercial Antigen Detection Enzyme-Linked Immunosorbent Assay (ELISA) using Non Structural protein 1 (NS1) (Bio Rad) for early definitive laboratory diagnosis of dengue infection under field conditions in Sri Lanka. A panel of acute serum samples collected from 99 patients clinically suspected of having dengue fever (<5 days) warded at the North Colombo Teaching Hospital, Ragama, Sri Lanka were used for the present study. Serum samples were tested using Antigen Detection ELISA according to the method described by the manufacturer. Results of this novel assay were compared with RT-PCR assay using Chi-squared test. Two variables were analyzed at a 95% confidence interval and P value <0.05 was considered as significant. Twenty two and 65 patients were positive and negative, respectively, for dengue infection by both assays. Nine patients were confirmed as dengue by the Antigen Detection ELISA only. Three patients were confirmed as dengue by RT-PCR assay only. Antigen detection ELISA showed 88% of agreement with the RT-PCR assay. According to the Chi-squared test, there was no significant difference between the two assays for early diagnosis of dengue infection (?2=46, P=0.0000). Novel commercial Antigen Detection ELISA kit (Bio-Rad 72830) can be used for early definitive laboratory diagnosis of dengue infection in Sri Lanka under field conditions. Acknowledgement: the International Atomic Energy Agency (SRL 06/28) for technical co-operation and APCOT Marketing LTD, Sri Lanka for supplying Antigen detection ELISA kits.