ICAR Research Data Repository for Knowledge Management
TaqMan real-time PCR assay based on DNA polymerase gene for rapid detection of Orf infection.
KRISHI: Publication and Data Inventory Repository
View Archive Info| Field | Value | |
| Title |
TaqMan real-time PCR assay based on DNA polymerase gene for rapid detection of Orf infection.
|
|
| Creator |
Bora DP, Venkatesan G, Bhanuprakash V, Balamurugan V, Prabhu M, Siva Sankar MS, Yogisharadhya R.
|
|
| Subject |
TaqMan PCR, Orf Virus, Clinical samples
|
|
| Description |
Both conventional and real time PCR (rt-PCR) assays based on the amplification of a 103bp fragment from the DNA polymerase (DNA pol) gene (conserved, non-structural) of Orf virus (ORFV) were developed for detection and semi-quantitation of ORFV DNA from infected cell culture and clinical samples. The latter technique was based on TaqMan chemistry. The rt-PCR assay was specific and sensitive as it could detect as low as 3.5fg or 15 copies of ORFV genomic DNA. Both intra- (0.38-1.0%) and inter-assay (0.53-2.87%) variabilities of rt-PCR were within the acceptable range meaning the high efficiency and reproducibility of the assay. The rt-PCR was applied successfully to detect ORFV DNA from suspected clinical samples. Further, the assay has shown a relative diagnostic sensitivity and specificity of 100% and 93.5%, respectively, when compared to B2L gene based semi-nested PCR implying a wide potential of this rt-PCR for rapid field diagnosis of Orf in sheep and goats.
|
|
| Date |
2023-05-12T10:40:02Z
2023-05-12T10:40:02Z 2011-11-01 |
|
| Type |
Research Paper
|
|
| Identifier |
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/77245 |
|
| Language |
English
|
|
| Publisher |
Elsevier
|
|