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Development of a RT-PCR ELISA for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants and its evaluation on clinical samples

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Title Development of a RT-PCR ELISA for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants and its evaluation on clinical samples
Not Available
 
Creator Dubey, P.
Mishra, N.
Rajukumar, K.
Behera, S.P.
Kalaiyarasu, S.
Nema, R.K.
Prakash, A.
 
Subject Pestivirus
RT-PCR
RT-PCR ELISA
Real-time RT-PCR
Ruminants; Virus isolation
 
Description Not Available
The aim of this study was to develop a reverse transcription polymerase chain reaction ELISA (RT-PCR ELISA) for detection of ruminant pestiviruses and to evaluate its diagnostic performance on clinical samples obtained from cattle, sheep and goats. Optimization was carried out by serial dilution of home-made digoxygenin-labelled RT-PCR product standards obtained from pestivirus isolates and pestivirus infected animals. The detection limit of the assay was 10TCID50/ml, similar to virus isolation and real-time RT-PCR but 10-fold higher than RT-PCR. The assay had high analytical specificity along with a good reproducibility. When the assay was evaluated on the samples obtained from animals infected experimentally with BVDV and from the field using virus isolation as standard, it showed a high diagnostic sensitivity (95.9%) and specificity (98.6%) and there was strong agreement (97.5% concordance) between the two tests. However, it displayed an increased diagnostic specificity and sensitivity over RT-PCR. Additionally, when a few samples (n=26) were tested by RT-PCR ELISA and real-time RT-PCR, 100% concordance was obtained between them. Our results showed that RT-PCR ELISA can be a rapid, cost effective and alternative molecular diagnostic test for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants in ordinary laboratory settings.
Not Available
 
Date 2018-03-22T07:50:46Z
2018-03-22T07:50:46Z
2015-03-01
 
Type Research Paper
 
Identifier Dubey, P., Mishra, N., Rajukumar, K., Behera, S.P., Kalaiyarasu, S., Nema, R.K., Prakash, A (2015) Development of a RT-PCR ELISA for simultaneous detection of BVDV-1, BVDV-2 and BDV in ruminants and its evaluation on clinical samples. J Virol Methods. 2015 Mar;213:50-6.
0166-0934
http://krishi.icar.gov.in/jspui/handle/123456789/5990
 
Language English
 
Relation Not Available;
 
Publisher Elsevier B.V.