SUMMARY Severe thrombocytopenia and increased vascular permeability are two major characteristics of dengue haemorrhagic fever (DHF). To develop a better understanding of the roles of platelet-associated IgG (PAIgG) and IgM (PAIgM) in inducing thrombocytopenia and its severity of disease in patients with secondary dengue virus infection, the relationship between the PAIgG or PAIgM levels and disease severity as well as thrombocytopenia was examined in 78 patients with acute phase secondary infection in a prospective hospital-based study. The decrease in platelet count during the acute phase recovered significantly during the convalescent phase. In contrast, the increased levels of PAIgG or PAIgM that occurred during the acute phase of these patients decreased significantly during the convalescent phase. An inverse correlation between platelet count and PAIgG or PAIgM levels was found in these patients. Anti-dengue virus IgG and IgM activity was found in platelet eluates from 10 patients in an acute phase of secondary infection. Increased levels of PAIgG or PAIgM were significantly higher in DHF than those in dengue fever (DF). An increased level of PAIgM was associated independently with the development of DHF, representing a possible predictor of DHF with a high specificity. Our present data suggest that platelet-associated immunoglobulins involving antidengue virus activity play a pivotal role in the induction of thrombocytopenia and the severity of the disease in secondary dengue virus infections.

Background Dengue cases have been continuously reported in Nepal, including some large outbreaks, since its first introduction in 2004. The disease is now expanding towards newer locations above 1400 m high, especially the country's capital city, Kathmandu. In 2019, >14,000 dengue cases including six deaths were reported. This study was aimed at the detection and molecular characterization of dengue virus (DENV) in dengue patients. Methods A total of 451 patients were enrolled in this study. Demographic, clinical and laboratory information was collected from dengue patients. Dengue infection was confirmed by antibody/antigen detection assays followed by RT-PCR analysis. Results The DENV patients showed fever, body ache, headache, myalgia, retro-orbital pain and arthralgia. The platelets were decreased, serum liver enzymes were increased and leucopenia was seen. Out of 195 patients, 111 (57.0%) were positive for DENV RNA by consensus PCR. We found DENV-2, 70 (63.1%) as the predominant serotype responsible for the 2019 outbreak, while DENV-3 was detected in two patients. Conclusion Our findings suggest that DENV-2 was the major serotype causing the 2019 massive outbreak in Nepal. This information will help in disease control programs to understand the molecular epidemiology and its changing trend.

Japanese encephalitis (JE) is a serious health problem in the southwestern region of Nepal. Serological diagnostic kits for routine diagnostic use in this region have not been available. This study was performed to examine if the particle agglutination (PA) assay for Japanese encephalitis virus (JEV) IgM could be applicable to the samples collected in Nepal and also to evaluate the accuracy of clinical diagnosis of JE. One hundred and ninety-three blood samples were collected from the patients clinically diagnosed with JE or other infectious diseases in the JE-endemic, southwestern region of Nepal, in 2000. The PA assay was performed on these 193 serum samples and the results were compared with those by IgM-capture ELISA. Eighty-six samples were IgM-positive by the PA assay, and 71 of 86 were also positive by IgM-capture ELISA (sensitivity, 99%; specificity, 88%; positive predictive value, 0.82; negative predictive value, 0.99). These results suggest that the PA assay is a simple, reliable and useful diagnostic test to support clinical diagnosis in rural hospitals of Asia including Nepal.

Background:Dengue is an emerging public health problem in Tarai Region of Nepal. The most affected are among the poorest populations living in remote, rural areas and urban slums who have even no access for medical treatment. This study was carried out with the objective of the determining the serodiagnosis of dengue in symptomatic individuals and comparing it to IgM ELISA. Methods:One hundred eighty three samples were collected from the suspected cases having clinical symptoms of dengue in the Tarai region of Nepal during September to October 2007. Particle Agglutination (PA) assays was performed and it was compared with IgM capture ELISA. Results:Out of the total of 183 serum samples that 55(30%) samples were positive by PA assay. When compared with IgM-capture ELISA, a 50 of 55 were positive (sensitivity of 98% and specificity of 96%; a positive prediction value of 90% and negative prediction value of 99%). Conclusions:The finding of this study showed that dengue is firmly established in low endemic Tarai region of Nepal. The sensitivity and specificity of PA assay is acceptably high and will be useful in rural areas of Nepal.

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