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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty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 A Mixed infection of Plasmodium falciparum and Plasmodium malariae: the first report of a Plasmodium malariae infection after 37 years of its absence in Sri Lanka(2008) Hapuarachchi, H.A.C.; Abeysundara, S.; Gunawardena, N.K.; Manamperi, A.; Senevirathne, M. P.; Leemingsawat, S.; Chavalitshewinkoon-petmitr, P.; de Silva, N.R.; Abeyewickreme, W.Malaria has been endemic in Sti Lanka for several centuries. Currently, only Plasmodium falciparum and P. vivax are present in the country. P. malariae infections have not .been reported in Sri Lanka since 1969. The objective is to determine the presence of malaria species in a patient returned from Malawi. The clinical history of intermittent high fever for 2 weeks accompanied by severe headache, myalgia, arthralgia, vomitimg, loss of appetite and backache with ictetus and mild hepatosplenomegaly suggested malaria in this 51 year old patient. Apart from the basic biochemical investigations, presence of malarial species was determined by light microscopy and confirmed by Real-Time Polymerase Chain Reaction (PCR) technology. Biochemical investigations showed a high serum bilirubin (4.8 mg/di) and liver enzyme (SGOT = >125 units, SGPT = >250 units) levels. Serum haemoglobin level (12.8 g%) was normal. Except for the presence of ptoteinuria (albumin = ++), bile (+) and red blood corpuscles (RBC) in his urine, renal functions were normal. Microscopical examination of Giemsa stained thin and thick blood smears showed an asexual parasite density of 120,000 per ul of blood. Infected RBCs were not enlarged, The presence of double-chromatin and applique form trophozoites, occasionally invading multiple RBCs suggested P. falciparum infection. In addition, there were characteristic band form trophozoites of P. malariae. Real-Time PCR protocol confirmed the presence of both P. falciparum and P. malariae in this patient. This is the first case of P. malariae reported in Sri Lanka after 4 decades, though the infection had been acquired from Malawi. Clinical and biochemical evidence indicated liver dysfunction and a transient glomerulonephritis, both of which subsided after treatment with quinine. This case report emphasizes the need of physicians to be more vigilant about the presence of malaria among immigrants, despite the drastic reduction of malaria in the country in recent years. Hence, this report highlights the importance of a proper programme in Sri Lanka to screen immigrants for infectious diseases.Item Clinical utility of PCR and real time PCR assays for Cytomegalovirus, hepatitis B and hepatitis C infections.(Sri Lanka Association for the Advancement of Science, 2008) Dassanayake, R.S.; de Silva, P.; Weerasena, J.; Gunawardene, Y.I.N.S.; Manamperi, A.Molecular Medicine Unit, Faculty of Medicine, University of Kelaniya, Reactivation of cytomegalovirus (CMV), Hepatitis B (HBV) and C (HCV) viruses from the status of latency is seen in immunocompromised individuals and such reactivation is often associated with morbidity and mortality in such individuals. The prevalence of these viral infections in a selected population of patients referred to the Molecular Diagnostic Laboratory at the Durdan's Hospital, Colombo, during the period from August 2007 to May 2008 were studied using qualitative PCR assays. All specimens from patients with suspected clinical diagnoses of either CMV or HBV or HCV infections were analyzed. Of 176 samples analyzed for CMV 78 were positive (37 males, 29 females) and majority of them are patients from a nephrology unit. Out of 40 and 10 samples analyzed from males and females, respectively, 22 and 4 were positive for HBV. Twenty six samples were analyzed for HCV and only 6 were fond to be infected with viruses and all of them were from males. Although PCR detection of these viral DNA/RNA is a sensitive method to detect infection, it lacks specificity for the detection of active viral disease and for monitoring the efficacy of antiviral therapy. Therefore, Real-time PCR (RT-PCR) assays for the detection and quantification of CMV-DNA, HBV-DNA and HCV-RNA were developed using SYBRgreen1 chemistry. The assays developed are capable of detecting viral particles in blood samples and quantifying viral DNA accurately over a broad range of input target copies (102 - 108copies/ml) and therefore, can be used to predict the reactivation of viruses by comparing with published kinetic criteria in clinical guidelines. Post PCR analyses of Real-time PCR products by agarose gel electrophoresis revealed bands having the same intensity for a wide range of target copies (103 -108copies/ml). In contrast, RT-PCR elicited higher cycle threshold for the descending order of concentration of target copies. Therefore, based on these results, it is evident that the intensity of conventional PCR bands should not be used for the assessment of viral reactivation or for monitoring therapeutic intervention and for this purpose RT-PCR is the method of choiceItem Rapid differential diagnosis of dengue and chikungunya infections by multiplex RT-PCR and impact of chikungunya infection on liver biochemical tests(Sri Lanka Association for the Advancement of Science, 2008) Manamperi, A.; de Silva, P.; Ekanayaka, C.; Gunawardene, Y.I.N.S.; de Silva, J.; Weerasena, O.V.D.S.J.; Dassanayake, R.S.Item Application of a Real Time Polymerase Chain Reaction (PCR) for Detection of Pathogenic Leptospira in Clinical Samples(University of Kelaniya, 2012) Denipitiya, D.T.H.; Jiffrey, A.M.; Abeyewickreme, W.; Wellawaththge, C.; Hapugoda, M.D.Leptospirosis, is a zoonotic disease with worldwide distribution, caused by pathogenic species of the genus Leptospira. It has the greatest impact on health in developing countries where it is often grossly under-recognized. Clinical features are similar to a range of other infectious diseases that occur in the same environmental and climatologic conditions. Therefore, laboratory confirmation is essential for proper management of leptospirosis patients. Molecular assays offer definitive laboratory confirmation of leptospirosis at the early phase of infection (1-5 days of fever) within a few hours. The objective of this study was to establish and evaluate potential use of a real time- PCR assay for early, definitive laboratory confirmation of leptospirosis patients. A SYBR green-based real time PCR assay targeting a 203 bp fragment on the secY gene which is conserved among pathogenic serovars of Leptospira was established using a reference DNA sample (Leptospira interrogans strain RGA). Analytical specificity of the assay was tested with the DNA from pathogenic and non-pathogenic Leptospira spp. and five other micro organisms. Analytical sensitivity of the assay was tested using serial dilutions of the reference sample. A panel of acute blood samples (n=150) collected during early phase of infection (1-5 days of fever) from leptospirosis suspected patients was used for evaluation of real time PCR vs qualitative PCR. The results show, real time PCR assay with high analytical specificity (100%) was established and the assay shows 100 times higher sensitivity over qualitative PCR assay (1.3 pg/ml). Real time PCR and qualitative PCR could diagnose current leptospirosis infection in 37.3% (56/150) and 19.3% (29/150) suspected patients respectively. These results indicate high sensitivity of real time PCR over qualitative PCR for diagnosis of leptospirosis patients. In conclusion, this study shows that real time PCR has the potential to facilitate rapid and sensitive diagnosis of acute leptospirosis during early phase of infection.