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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 Role of Aedes albopictus in transmitting dengue virus in some endemic areas in Kurunegala District.(University of Kelaniya, 2003) Hapugoda, M.D.; de Silva, N.R.; Abeysundara, S.; Bandara, K.B.A.T.; Dayanath, M.Y.D.; Abeyewickreme, W.Abstract AvailableItem Detection of dengue virus in Aedes albopictus mosquitoes by Reverse Transcription Polymerase-Chain Reaction-Liquid Hybridization (RT-PCR-LH) based assay.(Sri Lanka College of Microbiologists, 2003) Hapugoda, M.D.; Gunasekera, M.B.; de Silva, N.R.; Gunasena, S.; Prithimala, L.D.; Dayanath, M.Y.D.; Abeyewickreme, W.Dengue is an important public health problem. In this study an RT-PCR-LH assay was developed for the detection of dengue virus in Ae.albopictus, a vector of dengue. Laboratory bred Ae.albopictus (adults inoculated with dengue prototypes were tested by RT-PCR-LH assay. RT-PCR products of NS3 gene of 4 dengue prototypes were hybridized in liquid phase with 32P) labelled cocktail of dengue serotype-specific ologonucliotides. Semi-Nested-PCR agarose gel electrophoresis (Semi-Nested-PCR-AGE) assay with dengue type specific oligonucliotides was carried out for typing of RT-PCR products. Wild-caught Ae.albopictus (larvae (n=89 pools) and adults (n=69 pools) collected from dengue case reported stations during the period of 1999-2002 were also tested by RT-PCR-LH and typed by Nested-PCR-AGE assay). A DNA band (470bp) specific for dengue virus was observed in all pools of Ae.albopictus (inoculated with dengue prototypes in RT-PCR-LH assay. When RT-PCR products of dengue prototypes inoculated mosquitoes were typed by Semi-Nested-PCR-AGE assay, bands of 169,362, 265, 426 bp sizes corresponding to DEN1, DEN2, DEN3 and DEN4 respectively were observed. The DNA band specific for dengue virus (470bp) was also observed in 6 pools of wild-caught adults in RT-PCR-LH assay. They were found to be infected with DEN3 (265bp DEN3 specific DNA band was detected) by Semi-Nested-PCR-AGE assay. None of the wild-caught larvae showed dengue specific DNA band (470bp) in RT-PCR-LH assay). RT-PCR-LH with Semi-Nested-PCR-AGE assays are useful for the detection and typing of dengue virus in Ae.albopictus. Ae.albopictus (in Sri Lanka is competent in transmitting DEN3 and possibly other serotypes. Detection of dengue virus for the first time in Ae.albopictus in Sri Lanka confirms earlier observations that it may play an important role in transmitting dengue). Acknowledgements: Financial assistance by the International Atomic Energy Agency (Technical Co¬operation grant no SLR/ 06 / 024) and University of Kelaniya (Research grant no RP/03/04/06/01/00) is gratefully acknowledged.Item Silent transmission as a risk factor affecting transmission of dengue fever.(Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, 2007) Hapangama, H.A.D.C.; Hapugoda, M.D.; Gunawardene, Y.I.N.S.; Premaratna, R.; Dayanath, M.Y.D.; Abeyewickreme, W.The global incidence of dengue fever has increased by more than four-folds over the last 30 years, making it the most threatening mosquito-borne viral disease at present. Objective of this study is to determine the role of silent transmission •on' incidence of dengue. A total of 40 households, living within a 300 m radius of seven selected ‘confirmed dengue cases at different dengue high risk localities in Gampaha District were recruited for this study. A minimum of three inhabitants were tested for anti-dengue antibodies using a commercial kit to determine the prevalence of silent transmission on dengue infection in each households. Entomological surveillance was carried out in all seven localities. Out of 40 households, 26 (65%) houses were positive for dengue viral infection. Total of 148 inhabitants (68 Males); mean age; 35.9 years were enrolled. Of the 148, 41 (27.7%) had evidence of exposure to dengue virus [positive for IgM: 28/41(68.4%), IgM'& IgG: 7/41(17%) and IgG: 6/41(14.6%)]. Out of 28 primary infections, 20(71.4%) were asymptomatic. Of the 7 secondary infections, 1(14.28%) was asymptomatic. Of the 6 previous exposures to dengue 4(66.67%) were asymptomatic. Of the 7 localities investigated, 1 had >50%, 4 had >25% and 2 had <25% clustering of cases. Ae. albopictus found in all seven localities and Ae. aegypti found only in two localities. This study suggests presence of silent transmission of dengue virus with a trend towards clustering around cases and also this shows that the presence of vectors increases the incidence of dengue.Item Transmission study of dengue fever in Gampaha District, Sri Lanka(Faculty of Tropical Medicine, Mahidol University, 2007) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Dayanath, M.Y.D.; Abeyewickreme, W.RATIONALE: Dengue is an important flaviviral infection in Sri Lanka.OBJECTIVES; To determine the effect of possible contributing factors on transmission of Dengue METHODS: A total of 40 households situated around 7 selected dengue cases confirmed by both IgM-IgG ELISA and HAI assay were taken. A minimum of 3 inhabitants from each household were tested for anti-dengue antibodies. At each household, mosquito surveillance was carried out. RESULTS: Of the 40 households, 26(65%) were positive for dengue viral infection. Of the 148 inhabitants, 41 had evidence of exposure to dengue. Of the 41. asymptomatic dengue infection was observed in 25(61%). Paired sera were collected from 36/41 volunteers and tested by HAI assay which confirmed dengue virus infection in 4(11.1%) and were confirmed secondary. Three (75%) of them were asymptomatic. Balance 32(88.9%) was diagnosed as recent dengue infections. Of that 21(65.6%) were asymptomatic probable secondary 7(33.3%), recent dengue 14(66.6%)]. Clustering of cases was observed in all localities varying from 56.5%. There was a significant statistical association between the presence of a definitive dengue infection with the presence of Aedes vectors (x2=3.1, p=0.1). Ae. albopictus were found in all localities 'while Ae. aegypti was found only in one. Breteau index for Ae. albopictus varied from 0-260 while- container index varied from 0-84%. Highest clustering of dengue cases were observed in localities with the highest indices for' mosquitoes. CONCLUSIONS: Study confirms that the presence of asymptomatic infections and clustering of cases. The predominant vector was Ae. albopictus.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 Effect of silent transmission and clustering of cases on transmission of dengue in Gampaha district(Sri Lanka Association for the Advancement of Science, 2007) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Premaratna, R.; Dayanath, M.Y.D.; Abeyewickreme, W.Silent transmission of dengue virus and clustering of cases have been suggested as possible factors for the increasing incidence of dengue fever. Objective of this study was to determine the presence of silent transmission and clustering of cases of dengue fever in the Gampaha District. Study was carried out using cluster investigation method. A cluster consists of family members and immediate neighbours (minimum of 20) of a dengue index-case. Serum samples from volunteers were tested for anti-dengue antibodies using Dengue-Duo-IgM/IgG Rapid Cassette (Panbio diagnostics, Australia). Using 7 index cases, 148 volunteers (68 Males); mean age: 35.9 years were enrolled. Of the 148, 41 had evidence of exposure to dengue virus [positive for IgM: 68.4% (28/41), IgM & IgG: 17% (7/41) and IgG: 14.6% (6/41)]. Out of 28 primary infections, 71.4% (20/28) were asymptomatic. Of the 7 secondary infections, 14.28% (1/7) was asymptomatic. Of the 6 previous exposures to dengue, 4 (66.67%) were asymptomatic. There was no significant association between sex and exposure to infection [31% (21/68) males vs 25% (20/80) females, p>0.05] or between sex and occurrence of symptoms among exposed individuals [71% (15/21) males vs 50% (10/20) females, p>0.05]. Older individuals aged over 40 years, were most likely to be asymptomatic than younger persons (94% (14/15 exposed) vs 50% (13/26 exposed), P<0.01). Out of 7 clusters investigated, 1 had >50%, 4 had >25% and 2 had <25% clustering effects. A high proportion of asymptomatic infections were observed among adults over 40 years without gender difference. Study suggests persistence of silent transmission of dengue virus with a trend towards clustering around cases. Acknowledgement: World Health Organization (WHO/SEARO SN1144) and technical co-operation by International Atomic Energy Agency (TC/SRL 06/28)