Clinical Investigation
Prevalence and Molecular Characterization of Dengue Viruses Serotypes in 2010 Epidemic

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Abstract

During August to November 2010, a dengue (DEN) outbreak occurred, and several cities of Pakistan were affected. In this study, serum samples from 317 patients, based on clinical symptoms and blood test, were included from Lahore, Sheikhpura and Gujranwala cities for further diagnosis of different genotypes of DEN virus. The samples were categorized into 2 groups: group 1 containing patients with dengue fever (DF) without hemorrhagic manifestations (DF) (n=228) and group 2 involving patients having DF with hemorrhagic manifestations (dengue hemorrhagic fever) (n=89). The samples found positive for DEN in reverse transcriptase polymerase chain reaction (PCR) by targeting capsid and premembrane genes containing fragment of size ~2.5 kb for groups 1 and 2 were found to be 83.33% and 91.01%, respectively; however, the detection limit was quite low by immunoglobulin M antibody capture enzyme linked immunosorbent assay in case of both groups and was found to be 34.70% and 68.53%, respectively. The 16.67% and 8.99% samples of groups 1 and 2, respectively, were found negative in reverse transcriptase PCR analysis. In case of group 1 and group 2, by adopting seminested multiplex PCR strategy, predominant genotype was DEN virus type 2 with prevalence rates of 45.17% and 65.16%, respectively. DEN virus type 1 was the second most prevailing genotype in the population with prevalence rates of 18.85% and 19.10% for groups 1 and 2, respectively.

Section snippets

Sample Collection

In the current clinical study, a total of 317 serum samples were obtained from suspected individuals of Lahore, Sheikhpura and Gujranwala cities of Pakistan between August and November 2010 showing symptoms of headache, fever, rash, bleeding, myalgia and arthralgia and consistent temperature above 38°C. The patients were divided into 2 categories: patients having DF without hemorrhagic manifestations (n=228) and patients having DF with hemorrhagic manifestations based on clinical symptoms

RESULTS

In this study, a total of 317 serum samples were included and divided into 2 main groups: patients having DF without hemorrhagic manifestations (DF) (n=228) and patients having DF with hemorrhagic manifestation symptoms (DHF) (n=89) and evaluated further for parameters as described in Table 1. Fever (temperature > 38°C), headache, myalgia and decreased platelet counts were observed in almost all cases; however, these conditions were more severe in patients having DF with hemorrhagic

DISCUSSION

A total of 317 patients were included in this study and divided into 2 groups: patients having DF without hemorrhagic manifestations (DF) (n=228) and patients having DF with hemorrhagic manifestation symptoms (DHF) (n=89). The patients were initially diagnosed by general clinical symptoms and blood analysis, which include platelet count, leukocyte count and hematocrit percentage, which were disturbed frequently in most of the cases in DF and DHF groups.

The patients of both DF and DHF categories

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