Browsing by Author "Handunnetti, S.M."

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ABO-blood-group types and protection against severe, Plasmodium falciparum malaria
(Academic Press, 2005) Pathirana, S.L.; Alles, H.K.; Bandara, S.; Phone-Kyaw, M.; Perera, M.K.; Wickremasinghe, A.R.; Mendis, K.N.; Handunnetti, S.M.
Changes in full blood count parameters in leptospirosis: a prospective study
(BioMed Central, 2014) de Silva, N.L.; Niloofa, M.; Fernando, N.; Karunanayake, L.; Rodrigo, C.; de Silva, H.J.; Premawansa, S.; Handunnetti, S.M.; Rajapakse, S.
BACKGROUND: Leptospirosis presents diagnostic challenges to clinicians, in settings where other acute febrile illness are prevalent. The patterns of serial changes in haematological parameters in leptospirosis has not been evaluated previously. METHODS: Clinical and laboratory data were collected prospectively from patients with leptospirosis in two hospitals in Sri Lanka. Leptospirosis was diagnosed based on WHO clinical criteria with confirmation using Microscopic Agglutination Test titre > 400 or 4 fold rise between acute and convalescent samples. Full blood count parameters were analysed up to the 14th day of illness. RESULTS: Data from 201 patients with leptospirosis were available. Leukocyte counts and absolute neutrophil counts showed a decline over the first 5 days of illness, then rose until the end of the second week. On day 3 of fever, the majority (75%) had normal leukocyte counts, and by day 5, leukocytosis was seen only in 38.1%; leucopenia was an uncommon finding. Lymphopenia was seen in over half on day 5, declining to just under a quarter of patients by day 10. Platelets declined over the first 6 days and then gradually rose. Thrombocytopenia was seen in nearly three-fourths of patients by day 5. Haemoglobin and haematocrit levels declined over the course of illness. Total white cell and neutrophil counts were higher, and haemoglobin and haematorcrit were significantly lower, in patients with severe disease. CONCLUSIONS: Neither leukocytosis nor lymphopenia were prominent features, while thrombocytopenia was seen during the 3rd to 5th day of illness, with dropping haemoglobin levels. Neutrophilia and low haemoglobin levels appear to predict severe disease. These findings may be of use to clinicians in differentiating leptospirosis from other acute infections like dengue, and could help in predicting severe leptospirosis.
Comparison of HPLC profiles of venom of Apis darsata fabricius (Giant Asian Honey Bee) and Apis mellifera Linnaeus (Western Honey bee)
(2016) Gunasekara, D.L.P.E.; Handunnetti, S.M.; Premawansa, S.; Dias, R.K.S.; Witharana, E.W.R.A.; Dassanayake, W.M.D.K.; Premakumara, G.A.K.; De Silva, N.R.
Diagnosis of Vespa affinis venom allergy:use of immunochemical methods and a passive basophil activation test
(Allergy, Asthma & Clinical Immunology, 2019) Gunasekara, P.; Handunnetti, S.M.; Premawansa, S.; Kaluarachchi, P.; Karunatilake, C.; Ratnayake, I.P.; Dias, R. K. S.; Premakumara, G. A. S.; Dasanayake, W. M. D. K.; Seneviratne, S.L.; de Silva, R.
Background: Allergy to Vespa affinis venom is common in the Asia Pacific region. Venom preparations for diagnosis are not commercially available for this species. Methods: The prominent allergens in V. affinis venom were identifiedusing immunochemical methods. Use of ImmunoCAP of Vespula vulgaris crude venom/its components and a passive basophil activation test (BAT) in the diagnosis of patients who had anaphylaxis to V. affinis venom (n = 30) were also accessed. The IgE double-positivity rates (positive to both hornet and honeybee) in ImmunoCAP and the passive BAT were determined. Results: High IgE reactivity was seen with the five allergens in V. affinis venom; 96% (29/30) for 34 and 24 kDa, 93% (28/30) for 45 kDa and 90% (27/30) reactivity for the 100 and 80 kDa respectively. IgE cross-reactivity was low with ImmunoCAP using V. vulgaris venom (43%; 13/30) and Ves v1 (3%; 1/30), but relatively high with Ves v5 (73%; 22/30). All patients (100%) were positive to V. affinis venom in passive BAT. In ImmunoCAP, a high double-positivity rate (76%; 23/30) was detected while no double-positivity was detected in passive BAT. Conclusions: High IgE reactivity for five allergens of V. affinis points to the potential of using these allergens in component resolved diagnosis (CRD). The passive BAT has shown its importance as a promising diagnostic tool with high accuracy. It would be particularly useful in cases with doubtful double-positive results of other diagnostic tests.
A Double antibody sandwich ELISA for the diagnosis of vivax malaria: a tool for further research
(University of Colombo, 2000) Seneviratna, G.D.C.N.; Manamperi, A.A.P.S.; Kapilananda, G.M.G.; Longacre, S.; Handunnetti, S.M.; Udagama-Randeniya, P.V.
A diagnostic assay for Plasmodium vivax based on a double antibody sandwich enzyme-linked immunosorbent assay (DAS ELISA) was established to detect the merozoite surface protein 1 (PvMSP1) in wild isolates. This assay was based on the recombinant protein p19, a C-terminal processing product of PvMSP1. Of the two anti-P. vivax monoclonal antibodies (MAbs) selected, A21 was used as the capture antibody while horse radish peroxidase labelled A8 served as the probing second antibody. Optimized conditions established for p19 based DAS ELISA with the exception of a lower concentration of Tween-20 in buffers were suitable to screen lysed whole blood of malaria patients. This assay had a specificity of 100 percent for P. vivax and all the isolates of P. falciparum tested negative. Of the P. vivax-infected blood samples, only those containing both compact and schizont stages were diagnosed positive. The rest of the isolates tested negative either due to stage specificity of this assay or to the antigenic diversity of PvMSP1 in wild isolates. This test demonstrated a sensitivity of 27.58 percent and an accuracy of 63.15 percent. The positive and negative predicted values of this ELISA were 100 percent and 57.14 percent, respectively. Therefore, the P. vivax specific DAS ELISA developed and tested in the present study is not sufficiently sensitive to be used as a diagnostic tool for vivax malaria. Nevertheless, a baseline was established for development of diagnostic ELISA for future use with a more appropriate antigen.
Evaluation of two immunodiagnostic tests for early rapid diagnosis of leptospirosis in Sri Lanka: a preliminary study
(BioMed Central, 2015) Eugene, E.J.; Handunnetti, S.M.; Wickramasinghe, S.A.; Kalugalage, T.L.; Rodrigo, C.; Wickremesinghe, H.; Dikmadugoda, N.; Somaratne, P.; de Silva, H.J.; Rajapakse, S.
BACKGROUND: Leptospirosis is often treated based on clinical diagnosis. There is a need for rapid laboratory diagnosis for this condition. The aim of this study was to compare the diagnostic accuracy of two rapid IgM based immunodiagnostic assays with the microscopic agglutination test (MAT), in acute leptospirosis infection. METHODS: MAT, IgM based immunochromatographic test (Leptocheck-WB) and IgM ELISA were performed using acute sera of patients clinically suspected to have leptospirosis (n = 83). Bayesian latent class modeling was used to compare the accuracy of these tests. RESULTS: Percentage positivity for MAT, Leptocheck-WB, and IgM ELISA were 48.1, 55.3, and 45.7 % respectively. Bayesian latent class modeling showed a combined positivity rate of leptospirosis of 44.7 %. The sensitivity of MAT, Leptocheck-WB and IgM ELISA were 91.4, 95 and 81.1 %, and specificity were 86.7, 76.4 and 83.1 %, respectively. CONCLUSIONS: Leptocheck-WB has high sensitivity, and, because it is quick and easy to perform, would be a good screening test for acute leptospirosis infection. IgM ELISA has good specificity, and is comparable with MAT; given that it is easier to perform and more widely available than MAT, it would be a more appropriate confirmatory test for use in hospitals with limited access to a specialized laboratory.
High levels of serum angiopoietin 2 and angiopoietin 2/1 ratio at the critical stage of Dengue Hemorrhagic Fever in patients and association with clinical and biochemical parameters
(American Society for Microbiology., 2020) Mapalagamage, M.; Handunnetti, S.M.; Wickremasinghe, A.R.; Premawansa, G.; Thillainathan, S.; Fernando, T.; Kanapathippillai, K.; de Silva, A.D.; Premawansa, S.
ABSTRACT:Longitudinal changes of serum angiopoietin 1 (Ang-1) and angiopoietin 2 (Ang-2) associated with endothelial stability in dengue patients with different disease stages were studied. Serum Ang-1 and Ang-2 levels were measured in confirmed dengue fever (DF) patients on admission (DFA, n = 40) and discharge (DFD, n = 20); in dengue hemorrhagic fever (DHF) patients on admission (DHFA, n = 40), at critical stage (DHFC, n = 36), and on discharge (DHFD, n = 20); and in healthy controls (HC, n = 25). DHFC had the highest serum Ang-2 and lowest Ang-1 levels compared to DFA, DHFA, and HC (P < 0.050). The ratio of serum Ang-2/Ang-1 in DHFC was the highest among all study categories tested (P < 0.001). Significant positive correlations were observed between serum Ang-1 and platelet count in DHFA (Pearson r = 0.653, P < 0.001) and between Ang-1 and pulse pressure in DHFC (r = 0.636, P = 0.001). Using a cutoff value of 1.01 for the Ang-2/Ang-1 ratio for DHFC, a sensitivity of 83.2% and a specificity of 81.2% discerning DF from DHF were obtained. Therefore, serum Ang-2/Ang-1 could be used as a biomarker for endothelial dysfunction in severe dengue at the critical stage. KEYWORDS: angiopoietin; biomarker; dengue fever; dengue hemorrhagic fever; endothelial dysfunction.
Hypervariability in a leading P.vivax malaria vaccine candidate, C-terminal merozoite surface protein 1
(Sri Lanka Association for the Advancement of Science, 2000) Manamperi, A.; Holm, I.; Perera, L.; Handunnetti, S.M.; Longacre, S.
It is widely accepted that the C-terminal 42 kDa (p42) and 19 kDa (p19) processing fragments of plasmodium Merozoite Surface Protein-1 (MSP-1) are targets of immune protection. To begin to assess the degree of polymorphism in these MSP-1 vaccine candidates, we have investigated the sequence diversity in the p.vivax MSP-1 p42 processing fragment, in 19 natural isolates, from p.vivax infected patients in Kataragama. Sequence analysis of PvMSP-1 p42 in the 19 PCR positive isolates reveald 11 sequences of Belem origin and 8 sequences of Salvador-1 (Sal-1) origin. Among the isolates, these two stains are 98-100% homologous across this region, with one notable exception. This corresponds to a highly polymorphic block of 38 amino acids (24% amino acid homology among isolates). However, this polymorphism appears to be derived largely by re-assorting a dimorphism at each variable position. This type of restricte variability suggests that in spite of its diversity, there may nevertheless be a defined structure for this region of the molecule and that the diversity may be functionally important. Alternatively, it may be specifically designed for maximal effect in immune evasion, as a highly exposed immunogenic loop structure. In striking contrast, a single nucleotide substitution was detected in the cysteine rich C-terminal 19 kDa region, resulting in a lysine to glutamate substitution. This was detected in only one isolate among the 19 isolates investigated for sequence diversity. Since the PvMSP-1 C-terminal antigen is clearly hypervariable in the context of natural infections, a vaccine based on a single version of this antigen, might not induce an effective immunity against the multiple forms. In contrast, the PvMSP-1 p19 domain appears to be well conserved and thus appears to be a considerably more promising vaccine candidate.
Immunochemical characterization of venom of Apis dorsata Fabricus (Bambara) in Sri Lanka (Hymenoptera; Apidae)
(Institute of Biochemistry, Molecular Biology and Biotechnology, University of Colombo, Sri Lanka., 2015) Gunasekara, D.L.P.E.; De Silva, N.R.; Handunnetti, S.M.; Dias, R.K.S.; Witharana, E.W.R.A.; Dasanayake, W.M.D.K.; Premawansa, S.
Immunochemical characterization of venom of medically important ants in Sri Lanka and determination of ant cross-reactivity with western ant species
(2016) de Silva, B.D.; Handunnetti, S.M.; Premawansa, S.; Dias, R.K.S.; De Silva, N.R.
Leptospirosis: challenges in diagnosis, and predictors of severity
(Ceylon College of Physicians, 2016) Rajapakse, S.; Fernando, N.; Niloofa, M.J.R.; de Silva, H.J.; Karunanayake, L.; Premawansa, S.; Handunnetti, S.M.
No Abstract available
A pilot study on comparison of rapid immunodiagnostics for confirmation of leptospirosis
(Sri Lanka Medical Association, 2012) Eugene, E.J.; Wickramasinghe, S.A.; Kalugalage, T.L.; Rodrigo, C.; Wickremesinghe, H.; Dikmadugoda, N.; Somaratne, P.; de Silva, H.J.; Rajapakse, S.; Handunnetti, S.M.
INTRODUCTION: In Sri Lanka, leptospirosis is mostly diagnosed on clinical grounds. Serological confirmation is not obtainable during the acute stage of the illness. There is a need for rapid immunodiagnostics for confirmation of leptospirosis. Two immunodiagnostic assays, ie: enzyme linkedimmnnosorbent assay (ELISA) and immunochromatographic technique Leptocheck-WB test (LCT) areused to detect leptospira specific IgM antibodies which are prevalent in early stages of acute infections. AIMS: To compare the efficacy of these two rapid immunodiagnostic assays with the microscopic agglutination assay (MAT) to determine their applicability. Methods: A set of sera (n=83) collected in 2010 for which MAT titres were available was used to perform IgMELISAandLCT. RESULTS: Positivity for LCT and IgM ELISA were 55.4% and 48.2% respectively, and both assays detected acute infection by day 3 of the illness. MAT> 400 was used as the reference standard. For LCT, the overall sensitivity, specificity, accuracy, PPV and NPV (86.5%, 75.0%, 79.6%, 69.6% and 89.4% respectively) were higher compared to the respective values for IgM ELISA (50.0%, 62.3%, 57.1%, 50.0%, 62.3%). The highest of these values were observed during the first week for LCT and during the second week for IgM ELISA. The highest agreement was observed between LCT and MAT>400 (p=0.568) and there was a good agreement between LCT and IgM ELISA (p=0.520). CONCLUSIONS: The high sensitivity and specificity, ease of use, and non-requirement of specialized skills and equipment makes LCT a good choice for screening, while IgM ELISA is an appropriate test for confirming acute leptopsirosis.
Protein Carbonyl as a biomarker of oxidative stress in severe Leptospirosis, and its usefulness in differentiating Leptospirosis from Dengue Infections
(Public Library of Science, 2016) Fernando, N.; Wickremesinghe, S.; Niloofa, R.; Rodrigo, C.; Karunanayake, L.; de Silva, H.J.; Wickremasinghe, A.R.; Premawansa, S.; Rajapakse, S.; Handunnetti, S.M.
Pathogenesis of disease severity in leptospirosis is not clearly understood whether it is due to direct damage by pathogen or by adverse immune responses. Knowledge on biomarkers of oxidative stress which could be used in identifying patients with severe illness has shown to be of great value in disease management. Thus, the main aim of this study was to assess the damage to serum proteins and lipids, and their significance as biomarkers of oxidative stress in severe leptospirosis. In regions endemic for both leptospirosis and dengue, leptospirosis cases are often misdiagnosed as dengue during dengue epidemics. Therefore, the second aim was to assess the potential of the oxidative stress markers in differentiating severe leptospirosis from critical phase dengue. We measured serum antioxidants (uric acid and bilirubin), total antioxidant capacity (AOC), protein carbonyl (PC) and lipid hydroperoxide (LP) in patients with severe leptospirosis (n = 60), mild leptospirosis (n = 50), dengue during the critical phase (n = 30) and in healthy subjects (n = 30). All patient groups had similar total antioxidant capacity levels. However, the presence of significantly high uric acid and total bilirubin levels may reflect the degree of renal and hepatic involvement seen in severe leptospirosis patients (p<0.02). Serum PC and LP levels were significantly higher in leptospirosis patients compared to critical phase dengue infections (p<0.005). Moreover, high serum PC levels appear to differentiate SL from DC [area under the curve (AUC) = 0.96; p<0.001]. Serum PC may be a reliable biomarker of oxidative damage to serum proteins to identify severe leptospirosis patients (AUC = 0.99) and also to differentiate severe leptospirosis from mild cases (AUC = 0.78; p<0.005) indicating its contribution to pathogenesis. Use of serum PC as an indicator of leptospirosis severity and as an oxidative stress biomarker in differentiating leptospirosis from dengue would provide the opportunity to save lives via prompt patient management.
Serum nitrite levels in Sri Lankan patients with leptospirosis
(Elsevier, Singapore, 2012) Gunaratna, R.I.; Handunnetti, S.M.; Bulathsinghalage, M.R.C.; Somaratne, P.; Jayanaga, A.; de Silva, H.J.; Rajapakse, S.
OBJECTIVE: To determine whether blood nitrite levels are elevated in patients with leptospirosis.METHODS: Male patients fulfilling clinical and epidemiological criteria for a diagnosis of leptospirosis were recruited. Those with MAT titre of ≤400 together with those seroconverting to a titer of ≤200 were included in the analysis. Serum nitrite levels were measured in these patients and age, sex matched healthy controls. RESULTS: Patients from 3 hospitals (n=75) were screened during a 3 month period from 28th June to 3rd September 2009, of whom 20 were eligible for the study. Serum nitrite levels were found to be significantly higher in patients with acute leptospirosis [n=20, (0.359±0.229)μ M] compared to controls [(n=13,(0.216±0.051)μ M](P=0.014). A significant correlation was also observed between the MAT titre and the day of illness (r = 0.547; P<0.0001). CONCLUSIONS: Serum nitrite levels are higher in patients with acute leptospirosis compared to age and sex matched controls. No correlation could be assessed with severity of illness, as sample size was inadequate to determine this