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Detecting SARS-CoV-2 using a simple real time molecular assay will be helpful for the mitigation efforts in low / middle income countries during the pandemic. We have developed and validated a rapid and simple real time loop mediated isothermal amplification assay (LAMP) for screening of SARS-CoV-2 infection in known infected and non-infected individuals.Six sets of primers were designed targeting the N-gene of the SARS-CoV-2 (Accession ID MN994468). LAMP reactions were performed using Warm Start 2X Master Mix and real-time PCR machine at 65 °C for 60 cycles with 15 s for each cycle. Results were read by visualizing turbidity under ultraviolet light and real time fluorescence detection through FAM channel of the real time PCR machine. We tested a total of 320 including 240 SARS CoV-2 positive (Ct values40) and 80 SARS CoV-2 negative samples as tested by a real time RT-PCR using the newly developed LAMP assay.A total of 206 out of 240 SARS CoV-2 positive samples were tested positive by the newly developed LAMP assay with a sensitivity of 86%. All 80 SARS CoV-2 negative samples were tested negative by the newly developed LAMP assay with a specificity of 100%.The newly developed real time LAMP assay has a sensitivity of 86% and specificity of 100% compared to the real time RT-PCR for the detection of SARS CoV-2. The new assay will be useful to screen large number of samples if adopted to minimize the time and cost.

BackgroundLack of point of care diagnostics is a major challenge for control of human leptospirosis. Immunoglobulin M enzyme linked immunosorbent assays (IgM ELISA) have been widely used for the diagnosis of leptospirosis. The purpose of the present study was to determine the validity of IgM ELISA in the diagnosis of leptospirosis in a Sri Lankan context. MethodsConfirmed cases of leptospirosis from the 2008 Sri Lankan outbreak of leptospirosis and a group of leptospirosis excluded febrile patients were selected for the validation study. Disease confirmation and exclusion was carried out using either paired sample MAT (optimized for the region) or qPCR or both. A commercially available IgM ELISA kit was used and the procedure performed according to the manufacturers’ instruction in the Department of Microbiology, Faculty of Medicine, University of Peradeniya. ResultsThe study sample included 88 confirmed cases of leptospirosis and a comparison group of 71 acute fever patients. Of the 88 confirmed cases selected, 53 reacted in IgM ELISA and of the comparison group, 38 gave a positive reaction. *MAT was carried out among acute and convalescent samples where the acute sample was obtained within the first 10 days of illness and the convalescent sample after the day 14. A regionally optimized MAT panel with 21 serovars were used for MATSensitivity and specificity of IgM ELISA, as a point of care diagnostic test for patients in this sample with acute leptospirosis, was 60.23% (95% CI 49.78, 69.82) and 46.48% (95% CI (35.36, 57.96) respectively. Diagnostic accuracy of the test was 54.09% (95% CI 46.34, 61.65 %). The ROC (Receiver-operator characteristic curve) curve for the IgM ELISA showed a value of .669 for area under the curve. Optimal cut off points were not detected due to the poor test parameters in this sample. ConclusionThis study shows the poor diagnostic capabilities of IgM ELISA during the acute phase of leptospirosis in high endemic settingsDOI: http://dx.doi.org/10.4038/sljid.v4i2.6952 Sri Lankan Journal of Infectious Diseases 2014; Vol.4(2):83-89

This is an open access article under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

This is an open access article under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

Dengue virus (DENV) infections are increasing with respect to incidence and severity in the Central Province of Sri Lanka. The objective of this study was to define the clinical, immunological, and virological profiles of patients admitted to the General Hospital, Kandy with clinically apparent dengue. Demographic, clinical, hematological parameters, liver enzymes (ALT and AST), and blood samples were collected from 292 patients with fever5 days post onset and fulfilling the WHO criteria for the diagnosis of dengue fever/dengue hemorrhagic fever (DF/DHF). Samples were analyzed for, anti-DENV IgM, IgG, and DENV nucleic acid. Myalgia was the commonest complaint by 65% of the patients. Packed cell volume was45% in 27% of the patients while 42.12% had reduced platelets and 62.67% had reduced white blood cell counts. In contrast to other studies, positive tourniquet test (PTT) and petechiae were not major indicators of DENV infection or severity of the disease. Clinical profiles were significantly different between DF and DHF/DSS and showed many similarities to that reported elsewhere. Altogether, 43 patients (14.73%) were viremic as detected by RT-PCR; 181 patients (62%) were positive for anti-DENV IgM, and 245 (84%) patients were positive for anti-DENV IgG. In combination, anti-DENV IgM and RT-PCR assays detected 224 (77.5%) of DENV infected cases, thus improving the DENV diagnosis rate. Hence, the diagnostic utility of PTT, anti-DENV IgM/IgG serology, or RT-PCR used alone in the early phase of illness is low in Sri Lanka but the diagnostic value can be improved by a combination of serology and RT-PCR. J. Med. Virol. 88:1703-1710, 2016. © 2016 Wiley Periodicals, Inc.

The objective of the present study was to assess the circulating TNF-α and IL-2 levels in dengue virus (DENV) infected patients and to correlate these with clinical severity of DENV infections. A single analyte quantitative immunoassay was used to detect TNF-α and IL-2 in 24 dengue fever (DF) and 43 dengue haemorrhagic fever (DHF) patients, 15 healthy adults and 6 typhoid patients. The mean TNF-α and IL-2 levels of DENV- infected patients were higher than that of healthy adults and typhoid patients. No significant difference in TNF-α levels was noted between DF and DHF patients (p=0.5) but a significant increase in IL-2 levels was observed in DHF compared with DF patients (mean of DF=59.7pg/mL, mean of DHF=166.9pg/mL; p=0.02). No significant association of TNF-α or IL-2 levels was noted with packed cell volume (>45), thrombocytopenia, leucopenia or the presence of viraemia. The liver function tests measuring AST (aspartate aminotransferase) (p=0.01) and ALT (alanine aminotransferase) (p=0.02) levels were significantly elevated in DENV-infected patients. AST:ALT was significantly elevated in DHF/DSS (dengue shock syndrome) compared with DF patients. A significant positive linear correlation was noted between AST and IL-2 (r=0.31; p=0.01) and ALT and IL-2 elevations (r=0.2; p=0.02). Thus, AST and ALT levels correlate with both disease severity and circulating IL-2 levels. We suggest a role for circulating IL-2 in liver dysfunction and propose that a combined assessment of AST/ALT in conjunction with IL-2 at the early stages of symptomatic DENV infection may be useful to predict the severe forms of dengue.

Dengue fever (DF) is a mosquito borne virus infection which is endemic to the tropical regions of the world. In Sri Lanka, the first sero-positive case was reported in the 1960s; since then the island has experienced several outbreaks of DF/dengue haemorrhagic fever (DHF). The disease is more prevalent in some parts of the country where rapid urbanisation has taken place. Diagnosis of DF/DHF is mainly done using the clinical features only due to the unavailability of laboratory diagnosis in many parts of the country and this might lead to over or under diagnosis of the disease. A rational diagnostic approach which combines the history and clinical profiles together with specific virological laboratory data would help in the correct identification of the disease. Furthermore, a feasible algorithm for the laboratory diagnosis of dengue will help to confirm the cases with a high level of clinical suspicion. This would then facilitate the notification of correctly identified cases to the public health authorities to assess the dengue burden. The scope of this review is to improve the existing laboratory diagnosis of DF/DHF by proposing a feasible algorithm to implement in Sri Lanka that would enable better detection of cases.

Global leptospirosis disease burden estimates are hampered by the lack of scientifically sound data from countries with probable high endemicity and limited diagnostic capacities. We describe the seroepidemiologic and clinical characteristics of the leptospirosis outbreak in 2008 in Sri Lanka. Definitive/presumptive case definitions proposed by the World Health Organization Leptospirosis Epidemiology Reference Group were used for case confirmation. Of the 404 possible cases, 155 were confirmed to have leptospirosis. Highest titers of patient seum samples reacted with serovars Pyrogenes (28.7%), Hardjo (18.8%), Javanica (11.5%), and Hebdomadis (11.5%). Sequencing of the 16S ribosomal DNA gene identified six infections: five with Leptospira interrogans and one with L. weilli. In this patient population, acute renal failure was the main complication (14.8%), followed by myocarditis (7.1%) and heart failure (3.9%). The case-fatality rate was 1.3%. This report strengthens the urgent need for increasing laboratory diagnostic capabilities to determine the causes of epidemic and endemic infectious diseases in Sri Lanka, a finding relevant to other tropical regions.

ObjectivesThe current study was undertaken to compare the sensitivity, specificity, positive predictive value and negative predictive values of a dengue IgM rapid immunochromatography (ICT) assay with a dengue IgM capture ELISA for the detection of anti-dengue virus IgM in patients clinically suspected of having dengue fever (DF). MethodsBlood samples (n=119) were collected from paediatric and adult patients suspected of having DF at Teaching Hospital, Peradeniya (THP) after 5 days of fever. The samples were analyzed for the presence of anti-dengue virus IgM using a commercial rapid dengue IgM ICT assay (Hexagon dengue, Human, Germany) and a commercial IgM capture ELISA (Panbio Diagnostics Inc, Australia) provided to THP by the Ministry of Health for the diagnosis of dengue fever. ResultsWhen tested using the rapid dengue IgM ICT assay, 80 blood samples were positive and 39 were negative for anti-dengue virus IgM. When tested using the dengue IgM ELISA, 100 sera were positive and 19 were negative for dengue IgM. Using the IgM capture ELISA as the comparator, the sensitivity and the specificity of the rapid assay were 79% and 94.7% respectively with a positive (PPV) and negative (NPV) predictive value of 98.8% and 46.2%. ConclusionIn comparison with the results of the IgM capture ELISA, the low sensitivity and NPV for anti-dengue virus IgM detection by the rapid dengue IgM ICT assay was noted. The low sensitivity and NPV means that patients with DF will be missed when using this test. In contrast, the high specificity and PPV indicate that this rapid assay is able to detect true positives. Since preliminary screening of DF is carried out widely using these rapid tests, the strengths and limitations of this and other similar tests require validation before use in diagnostic laboratoriesDOI: http://dx.doi.org/10.4038/sljid.v4i2.5925 Sri Lankan Journal of Infectious Diseases 2014; Vol.4(2):77-82