Jiming  Gao, Ning  Kong, Yihong  Xiao, Angke  Zhang, En-Min  Zhou, 
    Development of Real-Time PCR Methods for the Detection of CD163 and Porcine Reproductive and Respiratory Syndrome Virus <i>N</i> Genes in Marc-145 
  Cell,
    Journal of Animal and Veterinary Advances,
    Volume 11,Issue 23,
    2012,
    Pages 4489-4493,
    ISSN 1680-5593,
    javaa.2012.4489.4493,
    (https://makhillpublications.co/view-article.php?doi=javaa.2012.4489.4493)
    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) <I>N</I> genes in Marc-145 cells. Primers were designed based 
  on the sequence of highly conservative region of <I>PRRSV N</I>, <I>CD163</I> 
  and <I>&#946;-actin</I> 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 <I>PRRSV N</I> 
  gene positive plasmid and total RNA dilution end-point standard curve from Marc-145, 
  respectively. The minimum detection levels for <I>PRRSV N</I> gene, <I>CD163</I> 
  and <I>&#946;-actin</I> gene were 10 copies, 0.25 and 0.025 ng total RNA per 
  reaction mixture, respectively. The R<SUP>2</SUP> 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 <I>&#946;-actin</I> 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 <I>PRRSV N</I> and <I>CD163</I> 
  gene in Marc-145 cell line.
    Keywords: PRRSV;CD163;SYBR green I;quantitative real-time PCR;China