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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 Screening of hepatitis C (HCV) antibody reactive donors by RT-PCR in a sample population of blood bank donors in Sri Lanka(Malaysian Society of Parasitology and Tropical Medicine, 2007) Manamperi, A.; Gunawardena, N.K.; Nugawela, P.; Bandara, A.; Wellawaththage, C.; Bindusara, R.M.; de Silva, H.J.; Abeyewickreme, W.The practice of screening donors for HCV antibodies has substantially lowered the risk of acquiring HCV infection from a transfusion. However, detection of molecular markers in blood is the most reliable means of diagnosing active viral infection. Molecular studies on HCV antibody reactive donors have not been previously performed in Sri Lanka. The present study was carried out to investigate the RNA positive rates in a sample population of anti-HCV antibody reactive blood donors in Sri Lanka, with a view of determining whether RT-PCR testing for HCV RNA should be carried out at the initial donor screening level. Eighty nine (89) HCV antibody reactive donors were tested for the presence of HCV RNA by RT-PCR (sensitivity 200 copies/ml) during the period October 2005 to May 2006. The 89 serology positive donors were initially detected by a third generation ELISA by routine screening of an initial pool of 26,176 blood donors. Of the 89 Anti-HCV antibody positive donors (0.34% of the total donor pool), 6 (0.023% of the total donor pool, and 6.74 % of antibody positive individuals) were positive for HCV RNA. The prevalence of HCVRNA positivity was low in this cohort of Sri Lankan blood donors. This is in keeping with the low prevalence of HCV infection in the community. Routine individual HCV-RNA screening of donors does not seem cost-effective in our setting. The RNA negative, antibody positive profiles reflect either false positive serology results or donors who have been exposed to HCV previously and subsequently resolved their infections.Item Comparison of different RNA extraction methods for Dengue Reverse Transcription -Polymerase Chain Reaction (RT-PCR)(Sri Lanka Association for the Advancement of Science, 2011) Adihetty, D.D.; Wellawaththage, C.; Abeyewickreme, W.; Abeywickrema, K.; Hapugoda, M.D.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.Item Application of nucleic acid technology (NAT) in the diagnosis of active viral replication in HBV and HCV infections and evidence for HBV surface antigen mutants(Sri Lanka Association for the Advancement of Science, 2008) Manamperi, A.; Gunawardene, Y.I.N.S.; Hapuarachchi, C.; Bandara, A.; Wellawaththage, C.; Abeyewickreme, W.; de Silva, J.Introduction: The community prevalence of Hepatitis B (HBV) and hepatitis C (HCV) infections, although considered low (< 1%) in Sri Lanka based on serological markers, pose a significant health threat to patients in high risk groups. The early diagnosis of active viral infection is crucial in such situations to prevent further transmission and to enable the clinicians to initiate successful therapeutic interventions. Objective: This study was carried out to investigate the usefulness of polymerase chain reaction (PCR) in the diagnosis of active viral replication in HBV and HCV infections. Methodology: All specimens from patients with serological evidence of hepatitis B (HBV surface antigen and/or antibodies for HBV core protein) or hepatitis C (antibodies for hepatitis C core protein-Anti-HCV) and referred to the Molecular Medicine Unit from May 2005 to May 2008 were analyzed by PCR and reverse-transcription PCR (RT-PCR) for HBV DNA (n=130) and HCV RNA (n=95) respectively. Results: Of the 130 patients tested, 57 (44%) were positive for HBV DNA. The positive group of patients included 10 renal transplant patients, 4 multiply transfused patients, 4 paediatric patients with lymphoma, and 1 patient with cirrhosis. Six HBV DNA positive patients had negative HBsAg serology profiles indicating the possibility of surface antigen mutant strains. The HBV DNA negative patients with positive serology profiles indicate sero-converted/ patients with resolved infections or false positive serology results. Of the 95 patients tested, 14 (15%) were positive for HCV RNA and included 3 paediatric patients with thalassaemia. HCV RNA negative, anti-HCV positive profiles reflect either false positive serology results (due to less specific antibody assays) or donors who have been exposed to HCV previously and subsequently resolved their infections. Conclusions: A major proportion of patients with serological markers for HBV have active viral infection whereas only relatively a minor proportion of patients with serological markers for HCV have active viral replication. We have also found the first possible evidence of hepatitis B surface antigen mutant strains. This underlines the importance of the nucleic acid based technology in the diagnosis and assessment of infection with or suspected to have hepatitis B or C infections. We also emphasize the importance of introducing NAT for screening donors for HBV DNA and HCV RNA to substantially lower the risk of acquiring HBV/HCV infection from a transfusion.