Journal/Magazine Articles

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This collection contains original research articles, review articles and case reports published in local and international peer reviewed journals by the staff members of the Faculty of Medicine

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    Composition of malaria vectors and diversity of anopheline breeding habitats in the district of Mannar, Sri Lanka
    (Central Environmental Authority (CEA), 2016) Gunathilaka, P.A.D.H.N.; Hapugoda, M.D.; Wickremasinghe, A.R.; Abeyewickreme, W.
    In the malaria elimination phase in Sri Lanka, investigation on biological and ecological factors of malaria vectors are important in planning appropriate vector controlling strategies. Lack of sufficient biological and ecological information on malaria vectors in the Northern Province of the country, a malaria endemic region, is a major constrain in successful implementation of malaria control programmes. Therefore, the objective of this study was to explore the diversity of breeding habitats and species composition of malaria vector mosquitoes in the District of Mannar, Sri Lanka. Potential habitats for Anopheles mosquito larvae were surveyed from June, 2010 to July 20 J2 on a monthly basis in selected sampling sites in the Mannar District: Mannar Town, Vankalai and Silawathiura, within a radius about 20 km. In each site, 4 sub sites were selected A total of 37,788 Anopheles representing ten species was recorded from 12 breeding habitat categories. Built wells and waste water collections were conducive for anopheline breeding. Anopheles subpictus (96.2%, n= 36,351) was the dominant species followed by An. peditaeniatus (1.47%, n= 557), An. barbirostris (1.23%, n= 463), An. nigerriums (0.75%,n = 285), An. varuna (0.19%, n= 74), An. barbumbrosus (0.1%, n= 38), An. vagus' (0.03%, n= 12), An. pallidus (0.01%, n= 4), An.jamesii (0.05%, n= 2) and An. pseudojamesi (0.05%, n= 2). Use of wells and waste water drains as breeding places by potential malaria vectors indicates that both of these habitats act as larval reservoirs during the dry season. Presence of theses habitats in close proximity to human habitats create a potential risk of malaria transmission among humans. Therefore, health authorities need to be vigilant on these new habitats in vector control programmes.
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    Application of a real time Polymerase Chain Reaction (PCR) assay for the early diagnosis of human leptospirosis in Sri Lanka
    (Academic Press, Elsevier, 2016) Denipitiya, D.T.H.; Chandrasekharan, N.V.; Abeyewickreme, W.; Hartskeerl, C.M.; Hartskeerl, R.A.; Jiffrey, A.M.; Hapugoda, M.D.
    Leptospirosis has a major impact on health in Sri Lanka but is probably grossly under-recognized due to difficulties in clinical diagnosis and lack of diagnostic laboratory services. The objective of this study was to establish and evaluate a SYBR Green-based real-time Polymerase Chain Reaction (rt-PCR) assay for early, rapid and definitive laboratory diagnosis of leptospirosis in Sri Lanka. The rt-PCR assay was established and analytical specificity and sensitivity were determined using reference DNA samples. Evaluation of the assay for diagnosis of clinical samples was performed using two panels of serum samples obtained from 111 clinically suspected adult patients. Patients were confirmed as leptospirosis (n = 65) and non-leptospirosis (n = 30) by the Patoc - MAT. Other 16 samples gave ambiguous results. The analytical sensitivity of the rt-PCR was approximately 60 genome copies and no cross-reactivity was observed with saprophytic Leptospira spp. and other pathogenic microorganisms. Based on confirmation with Patoc-MAT on paired samples this corresponds to a diagnostic sensitivity and specificity of 67.7% (44/65) and 90.0% (27/30), respectively. This study showed that rt-PCR has the potential to facilitate rapid and definitive diagnosis of leptospirosis during early phase of infection in Sri Lanka.
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    A Comparative retrospective study of RT-PCR-based liquid hybridization assay for early, definitive diagnosis of dengue
    (Oxford University Press, 2010) Hapugoda, M.D.; de Silva, N.R.; Khan, B.; Dayanath, M.Y.D.; Gunesena, S.; Prithimala, L.; Abeyewickreme, W.
    Dengue is an important flaviviral infection in tropical and subtropical regions. Early diagnosis of dengue infection helps in monitoring the disease, determining when hospital admission is necessary and reducing case fatalities. The objective of this study was to carry out a retrospective comparison of an RT-PCR-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 four dengue type-specific DNA probes in liquid phase. The assay was validated in a comparative retrospective study using acute serum samples collected from 119 fever patients with or without dengue, confirmed by haemagglutination inhibition (HAI) assay, the gold standard assay for diagnosis of dengue infection. The RT-PCR-LH assay was highly specific for dengue and, as an early laboratory diagnostic method, had 100% sensitivity in detecting dengue patients confirmed by HAI assay. A high analytical sensitivity of two fluorescent focus units of dengue virus/reaction was achieved. This 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 when serological methods are of little value.
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    Re emergence of Chikungunya virus in South-east Asia: virological evidence from Sri Lanka and Singapore
    (Cambridge University Press, 2010) Hapuarachchi, H.C.; Bandara, K.B.A.T.; Sumanadasa, S.D.; Hapugoda, M.D.; Lai, Y.L.; Lee, K.S.; Tan, L.K.; Lin, R.T.; Ng, L.F.; Bucht, G.; Abeyewickreme, W.; Ng, L.
    Chikungunya fever swept across many South and South-east Asian countries, following extensive outbreaks in the Indian Ocean Islands in 2005. However, molecular epidemiological data to explain the recent spread and evolution of Chikungunya virus (CHIKV) in the Asian region are still limited. This study describes the genetic Characteristics and evolutionary relationships of CHIKV strains that emerged in Sri Lanka and Singapore during 2006-2008. The viruses isolated in Singapore also included those imported from the Maldives (n=1), India (n=2) and Malaysia (n=31). All analysed strains belonged to the East, Central and South African (ECSA) lineage and were evolutionarily more related to Indian than to Indian Ocean Islands strains. Unique genetic characteristics revealed five genetically distinct subpopulations of CHIKV in Sri Lanka and Singapore, which were likely to have emerged through multiple, independent introductions. The evolutionary network based on E1 gene sequences indicated the acquisition of an alanine to valine 226 substitution (E1-A226V) by virus strains of the Indian sublineage as a key evolutionary event that contributed to the transmission and spatial distribution of CHIKV in the region. The E1-A226V substitution was found in 95.7 % (133/139) of analysed isolates in 2008, highlighting the widespread establishment of mutated CHIKV strains in Sri Lanka, Singapore and Malaysia. As the E1-A226V substitution is known to enhance the transmissibility of CHIKV by Aedes albopictus mosquitoes, this observation has important implications for the design of vector control strategies to fight the virus in regions at risk of chikungunya fever.
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    Laboratory confirmation of dengue and chikungunya co-infection
    (Sri Lanka Medical Association, 2008) Hapuarachchi, H.A.C.; Bandara, K.B.A.T.; Hapugoda, M.D.; Williams, S.; Abeyewickreme, W.
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    Single antigen detects both immunoglobulin M (IgM) and IgG antibodies elicited by all four dengue virus serotypes
    (American Society for Microbiology, 2007) Hapugoda, M.D.; Batra, G.; Abeyewickreme, W.; Swaminathan, S.; Khanna, N.
    The resurgence of dengue (DEN) virus infections in the last few decades coupled with the lack of a preventive vaccine and specific antiviral drugs has jointly contributed to making this a significant global public health problem. Currently, symptomatic supportive treatment and fluid replacement therapy are the only means available to minimize DEN-induced mortality. As the clinical symptoms associated with DEN virus infections are indistinguishable from those of many other viral, bacterial, and parasitic infections, specific diagnostic tests assume critical importance in the unequivocal identification of DEN virus infections. We have designed a novel chimeric antigen based on envelope domain III (EDIII), a critical antigenic region of the major structural protein of DEN viruses. We fused EDIIIs corresponding to each of the four DEN virus serotypes using pentaglycyl linkers, over expressed the resultant tetravalent chimeric protein in Escherichia coli, and affinity purified it in high yields, obtaining approximately 30 mg protein of >95% purity per liter of culture. We show that this tetravalent antigen could specifically recognize anti-DEN virus antibodies of both the immunoglobulin M (IgM) and IgG classes. Using a large panel of IgM antibody capture-enzyme-linked immunosorbent assay- and hemagglutination inhibition-confirmed DEN virus-infected and uninfected patient sera (n = 289), we demonstrate that this tetravalent antigen can function as a diagnostic tool of high sensitivity and specificity.
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    Geographical Information System (GIS)based risk mapping and warning system for monitoring transmission of dengue in Kurunegala district
    (2004) Hapugoda, M.D.; Senarathne, H.; Chandrasena, U.A.; de Silva, N.R.; Rajamanthri, S.; Abeyewickreme, W.
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    A Novel reverse transcriptase-polymerase chain reaction based-liquid hybridisation(RT-PCR-LH) assay for early diagnosis of dengue infection
    (Sri Lanka Medical Association, 2003) Gunasekera, M.B.; Hapugoda, M.D.; Gunasena, S.; Subasinghe, S.A.S.C.; Bandara, K.B.A.T.; Khan, K.B.; Abeyewickreme, W.
    BACKGROUND: Early definitive laboratory diagnosis of dengue is difficult with the tests in routine use at present. OBJECTIVE: To develop a reverse transcriptase-polymerase chain reaction based liquid hybridisation (RT-PCR-LH) technique for the rapid and early diagnosis of dengue. RESEARCH DESIGN: RT-PCR products of the NS3 gene of dengue virus prototypes and of a few positive sera for dengue virus by culture, were allowed to hybridise in liquid phase with a mixture of dengue specific radio-labelled oligonucleotides. The products were separated by PAGE and visualised by autoradiography. 78 suspected dengue sera were also tested by RT-PCR-LH method, and by IgM-ELISA and HAI tests, for comparison. RESULTS: Two DNA bands (approximately equal to 470 bp and approximately equal to 455 bp) specific to dengue virus, were observed. RT-PCR-LH assay takes only 24 h. Of the 78 suspected dengue acute sera tested, 45/78 were positive by RT-PCR-LH, 31/78 were positive by IgM-ELISA, and 14/78 had a HAI titre > or = 2560. Duration of fever was known in 72 cases, and infection was detected by RT-PCR-LH in 11/22 of cases with < 5 d fever and by IgM-ELISA in 1/22. In cases with 5 to 15 d fever RT-PCR-LH and IgM-ELISA/HAI titre > or = 2560 detected infection in 30/50 and 27/50 respectively. The 10 sera which were negative by RT-PCR-LH, but were positive by either IgM-ELISA or HAI titre > or = 2560 were all > 5 d fever cases. RT-PCR-LH together with IgM-ELISA were capable of detecting dengue infection in 56/78 of the suspected cases. CONCLUSION: RT-PCR-LH assay developed in this study appears to have an advantage over other diagnostic techniques for the early detection of dengue.