@article{MAKHILLJAVA201211233957,
title = {Development of Real-Time PCR Methods for the Detection of CD163 and Porcine Reproductive and Respiratory Syndrome Virus N Genes in Marc-145
Cell},
journal = {Journal of Animal and Veterinary Advances},
volume = {11},
number = {23},
pages = {4489-4493},
year = {2012},
issn = {1680-5593},
doi = {javaa.2012.4489.4493},
url = {https://makhillpublications.co/view-article.php?issn=1680-5593&doi=javaa.2012.4489.4493},
author = {Jiming,Ning,Yihong,Angke and},
keywords = {PRRSV,CD163,SYBR green I,quantitative real-time PCR,China},
abstract = {The objective of this study was to develop an RNA-dependent
real-time reverse-transcriptase PCR (real-time RT-PCR) Method for the detection
of relative levels of CD163 and Porcine Reproductive and Respiratory Syndrome
Virus (PRRSV) N genes in Marc-145 cells. Primers were designed based
on the sequence of highly conservative region of PRRSV N, CD163
and β-actin gene as the reference gene. The specificity and sensitivity
of real-time RT-PCR Method were determined with the amplification signals Tm
peaks in positive controls and the standard curve generated from PRRSV N
gene positive plasmid and total RNA dilution end-point standard curve from Marc-145,
respectively. The minimum detection levels for PRRSV N gene, CD163
and β-actin gene were 10 copies, 0.25 and 0.025 ng total RNA per
reaction mixture, respectively. The R2 and efficiency of standard
curves were 0.983 and 102.166% for CD163, 1 and 101.453% for PRRSV N and 0.996
and 90.969% for β-actin genes. The developed real time RT-PCR Method
described in this report is more rapid, specific and sensitive than the conventional
RT-PCR for the detection of relative levels of PRRSV N and CD163
gene in Marc-145 cell line.}
}