Diagnosis of Dengue Viral Infection in Haryana by Reverse Transcriptase PCR

Author(s): Amit Kumar, H. N. Verma, Sushil Kumar, Jatinder Singh Chera and Sachinandan De*

Abstract: Background: Dengue is a common mosquito-borne viral disease in India with both endemic and outbreak forms. It causes hospitalization of about five hundred thousand people with a higher mortality rate. Thus, early and correct diagnosis is very crucial for treatment of Dengue fever. Methods: A total of 25 blood samples were obtained from patients within the age range of 5 years to 64 years, who were having symptoms of Dengue illness. The samples were obtained within the 7 days of onset of fever. All the blood samples that were subjected to NS1 ELISA for Dengue diagnosis afterwards they analysed with our RT-PCR test and both the methods were compared. The targeted product size was ~240 bp. Results: Among the 25 Dengue virus suspected samples tested, 22 were found positive with RT-PCR assay and 20 with NS1 ELISA (two were equivocal cases). The size of the PCR product obtained after amplification with specific primers was ~240 bp. The amplified product was also verified by restriction digestion with KpnI enzyme and the bands obtained were of sizes ~183bp and ~57bp. Sequencing of the 240 bp PCR fragment was also performed to confirm the result. Conclusions: RT-PCR based assay was found to be more specific and sensitive as compared to the ELISA assay.

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