Trend of Dengue Viral Infection and Circulating Serotypes in Tripura, North East India.

One of the major causes of morbidity and mortality in tropical and subtropical regions is Dengue viral infection. This virus belongs to family flaviviridae comprising of four antigenically distinct serotypes DENV 1 - 4. A small number of studies conducted in North Eastern (NE) Region of India reported Dengue cases in Assam, Meghalaya, Nagaland, Manipur and Arunachal Pradesh. However, no studies have been conducted in the state of Tripura, with regard to pattern of Dengue viral infection and its circulating serotypes. Therefore, this study was undertaken to identify the serotypes circulating in Tripura. MethodsPatients with acute febrile illness were tested for detecting Dengue viral infection by MAC ELISA and / or NS1 detection test at Viral Research and Diagnostic Laboratory (VRDL), of a tertiary care centre in Tripura for a period of 3 years. All NS1 positive samples were further tested for presence of viral RNA by Reverse Transcriptase –PCR (RT - PCR) and serotyping was done using serotype specific primers. ResultsA total of 2515 acute febrile cases seen over a period of 3 years from 2014 to 2017 was tested for Dengue virus infection by serology. Out of 2515 of cases, 405 cases tested for NS1 antigen, where 10.61 % (43 / 405) was NS1 positive. The remaining 2110 cases were tested for IgM antibody MAC ELISA and 15.68 % (331 / 2110) was MAC ELISA positive. Out of all NS1 antigen positive cases 34.88 % of PCR positive and serotype characterisation showed DENV - 1 was predominant serotype followed by DENV - 2 and DENV - 4 respectively. ConclusionsThere is a rising trend of Dengue virus infection in Tripura with circulation of multiple serotypes. Moreover, cocirculation of multiple serotypes is a risk to the emergence of recombinant strains and also heterotypic infection in the near future might lead to development of DHF and DSS. Hence, molecular characterization of circulating serotypes may be helpful in addressing the probabilities of Dengue outbreak and possibilities of complications.

Differential replicative fitness of the four dengue virus serotypes circulating in Colombia in human liver Huh7 cells

ABSTRACT Dengue has been a significant public health problem in Colombia since the simultaneous circulation of the four dengue virus serotypes. The replicative fitness of dengue is a biological feature important for virus evolution and contributes to elucidating the behavior of virus populations and viral pathogenesis. However, it has not yet been studied in Colombian isolates. This study aimed to compare the replicative fitness of the four dengue virus serotypes and understand the association between the serotypes, their in vitro infection ability, and their replication in target cells. We used three isolates of each DENV serotype to infect Huh-7 cells at an MOI of 0.5. The percentage of infected cells was evaluated by flow cytometry, cell viability was evaluated by MTT assay, and the pathogenicity index was calculated as a ratio of both parameters. The replicative fitness was measured by the number of viral genome copies produced using quantitative PCR and the production of infectious viral progeny was measured by plaque assay. We showed that Huh-7 cells were susceptible to infection with all the different strain isolates. Nevertheless, the biological characteristics, such as infectious ability and cell viability, were strain-dependent. We also found different degrees of pathogenicity between strains of the four serotypes, representative of the heterogeneity displayed in the circulating population. When we analyzed the replicative fitness using the mean values obtained from RT-qPCR and plaque assay for the different strains, we found serotype-dependent behavior. The highest mean values of replicative fitness were obtained for DENV-1 (log 4.9 PFU/ml) and DENV-4 (log 5.28 PFU/ml), followed by DENV-2 (log 3.9 PFU/ml) and DENV-3 (log 4.31 PFU/ml). The internal heterogeneity of the replicative fitness within each serotype could explain the simultaneous circulation of the four DENV serotypes in Colombia.

Identification of prevalent dengue serotypes by reverse transcriptase polymerase chain reaction and correlation with severity of dengue as per the recent World Health Organization classification (2009)

Context: A definite link between distinct dengue serotypes and severe clinical manifestations has not been established yet. The WHO classification (2009) of dengue is more competent in diagnosing severe cases compared to traditional (1997) classification. Aims: This study aimed to identify prevalent dengue serotypes and to correlate the severity of dengue with the dengue virus (DENV) serotypes in target population as per the recent WHO classification (2009). Settings and Design: A retrospective comparative observational study was conducted from 1st January 2015 to 31st December 2015. Subjects and Methods: We tested 242 dengue NS-1 antigen ELISA-positive cases for serotyping by dengue reverse transcriptase-polymerase chain reaction (RT-PCR). Severity of each dengue case confirmed by RT-PCR was determined as per the recent WHO classification (2009). Results: On the basis of RT-PCR, dengue infection was confirmed in 135 (55.78%) patients. DEN-3 was the most common serotype found in 71 (52.6%) patients, followed by DEN-2 serotype with 44 (32.6%) patients. Nearly 2.22% cases of DEN-2 and 2.96% cases of DEN-3 serotype were having dengue with warning signs. Severe dengue was found in 2.22% cases of DEN-2 and 5.18% cases of DEN-3 serotypes. Thrombocytopenia, haemorrhagic manifestations and atypical presentations were found most commonly in DEN-3 followed by DEN-2 serotype. Coinfection with more than one serotype was observed in our study, with the most common coinfection pattern being DEN-2 and DEN-3 serotypes. Conclusions: DENV-3 and DENV-2 serotypes are prevalent in the region and are associated with a more serious clinical profile than other serotypes.

Serotipos de dengue en México durante 2009 y 2010 / Dengue serotypes in México during 2009-2010

Introducción. El dengue en México es un problema prioritario de salud pública. Desde el 2008 el Departamento para la Vigilancia Epidemiológica y Virológica del InDRE implementó un nuevo algoritmo de diagnóstico del dengue, que utiliza la Red de Laboratorios Estatales de Salud Pública, para favorecer la representatividad geográfica, la oportunidad, la sensibilidad y la especificidad de la información que se obtiene. Métodos. La identificación de serotipos se realizó a partir de muestras positivas a la proteína NS1 por ensayo inmunoenzimático (ELISA). Las técnicas que se utilizaron fueron: aislamiento viral, PCR punto final y, desde 2009, RT-PCR en tiempo real (qRT-PCR). Resultados. En 2009 se analizaron 6,336 muestras; en 2,944 de éstas (46.6%) se identificó el serotipo DENV-1 que predominó sobre el serotipo DENV-2; el serotipo DENV-3 sólo se identificó en dos casos en Guerrero y el serotipo DENV-4 en un caso en Chiapas. En 2010 se analizaron 2,013 muestras. Se identificó algún serotipo en 1,607 muestras (79.88%) y, nuevamente, el serotipo DENV-1 predominó en todo el país. En Chiapas se identificaron los serotipos DENV-1, 2 y 4 y en Jalisco los serotipos DENV-1 y 3. Además, se identificó la circulación del serotipo DENV-3 en Guerrero y apareció el serotipo DENV-4 en San Luis Potosí. Conclusiones. Por la selección de muestras para vigilancia virológica de dengue mediante la positividad a la proteína NS1 y por la introducción de la técnica de qRT-PCR se optimizó la identificación de serotipos circulantes. La alta endemia, los brotes en nuevas regiones, el predominio del serotipo DENV-1 por varios años y la introducción lenta de otros serotipos, principalmente DENV-3, pueden favorecer la aparición de formas clínicas graves de dengue. La vigilancia epidemiológica inteligente del dengue brindará información para un mejor entendimiento de la enfermedad y promoverá acciones para su control y prevención.

Background. Dengue is a public health priority in Mexico. Since 2008, the dengue diagnostic algorithm for epidemiological and virological surveillance has been improved at InDRE and the public health laboratory network (RLESP) to optimize geographic representation, opportunity, sensitivity and specificity of the produced information. Methods. Dengue serotype identification is based on ELISA NS1 positive samples. Methods used are viral isolation, endpoint PCR and, since August 2009, real-time PCR (qRT-PCR). Results. In 2009, 6,336 serum samples were analyzed and 2,944 (46.6%) were positive for serotype identification. DENV-1 was detected in greater proportion followed by DENV-2, and DENV-3 4 was only identified in two cases in Guerrero and DENV-4 in one case in Chiapas. In 2010, 2,013 serum samples were analyzed and 1,607 (78.8%) were positive for serotype identification. DENV-1 was predominant throughout the country. In Chiapas, DENV-1, 2 and 4 were identified and in Jalisco DENV-1 and 3. DENV-3 was identified in Guerrero again and DENV-4 was detected in San Luis Potosí. Conclusions. The selection samples through NS1 positive samples and the introduction of qRT-PCR optimized serotype identification. Hyperendemicity, outbreaks in new geographic areas, the predominant circulation of DENV-1 for several years and the slow reintroduction of the other serotypes, mainly DENV-3, could increase clinical cases of severe dengue. An ¡intelligentí epidemiological surveillance program would offer information for a better understanding of the disease and promote action for its control and prevention.