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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.47 [2009] >
IJEB Vol.47(11) [November 2009] >
| Title: | Quantitative PCR: A quality control assay for estimation of viable virus content in live attenuated goat pox vaccine |
| Authors: | Kallesh, D J
Hosamani, M
Balamurugan, V
Bhanuprakash, V
Yadav, V
Singh, R K |
| Keywords: | Goat pox vaccine
PPR vaccine
TaqMan QPCR |
| Issue Date: | Nov-2009 |
| Publisher: | CSIR |
| Abstract: | Efficacy of live viral vaccine and
vaccine-induced sero-conversion depends on the optimum number of live virus
particles in a vaccine dose, which is one of the important aspects of quality
control. In the present study, TaqMan® probe quantitative polymerase chain reaction
(QPCR) based on conserved DNA pol gene of capripoxvirus was developed for the
quality control of attenuated monovalent goatpox and/or combined attenuated
goatpox and peste des petits ruminants (PPR) vaccines. Cells infected with
vaccine virus were harvested at critical time point and subjected to QPCR. A
critical time point for harvest of Vero cells infected with various log10
dilutions of reference virus was determined to be 36 h (highest slope 3.062),
by comparison of slopes of standard curves established with harvests at
different time intervals. The assay method was evaluated using different
batches of goatpox vaccine, and bivalent goatpox and PPR vaccine. The titers
estimated by QPCR and TCID50 method were comparable to each other. The QPCR
assay thus, could be used as an alternate method or supplementary tool for
estimation of live GTPV particles in monovalent goatpox or bivalent goatpox and
PPR vaccines. |
| Page(s): | 911-915 |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.47(11) [November 2009]
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