Home CONTACT Acta Virologica 2004 Acta Virologica Vol.48, p.249-255, 2004

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Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

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Acta Virologica Vol.48, p.249-255, 2004

Title: DEVELOPMENT OF A N GENE-BASED PCR-ELISA FOR DETECTION OF PESTE-DES-PETITS-RUMINANTS VIRUS IN CLINICAL SAMPLES
Author: P. Saravanan, R.P. Singh, V. Balamurugan, P. Dhar, B.P. Sreenivasa, D. Muthuchelvan, A. Sen, A.G. Aleyas, R.K. Singh, S.K Bandyopadhyay

Abstract: A highly sensitive N gene-based PCR-ELISA for the detection of Peste-des-petits-ruminants virus (PPRV) was developed. The RT-PCR yielded a digoxigenin (DIG)-labeled product of 336 bp comprising a sequence from PPRV N gene, which was then detected by ELISA. The assay could detect the viral RNA in PPRV-infected tissue culture fluids with a titer as low as 0.1 TCID50/ml. The assay is 10,000 times more sensitive than a classical RT-PCR combined with agarose gel electrophoresis. The assay could detect the virus in the clinical samples, which were negative by conventional sandwich ELISA (S-ELISA). The percentage positivity of the assay in detecting the virus in clinical samples was 66.2% compared to 48.6% for S-ELISA. The assay was more sensitive than S-ELISA also in detecting the virus in early as well as late phases of the disease. In addition, the assay could also be used for differential diagnosis of PPRV and Rinderpest virus (RPV).

Keywords: clinical samples; diagnosis, differential diagnosis; PCR-ELISA; peste-des-petits-ruminants; Peste-des-petits-ruminants virus; rinderpest; Rinderpest virus; RT-PCR; sandwich ELISA
Year: 2004, Volume: 48, Issue: Page From: 249, Page To: 255



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