title: Development of real-time PCR for quantitation of simian immunodeficiency virus 2-LTR circles.
abstract: Non-human primates infected with simian immunodeficiency virus (SIV) represent a robust model to evaluate pre-clinical efficacy of HIV-1 preventive strategies and to determine the size of reservoir.
  
  
  
  We developed a real-time qPCR assay to specifically quantify episomal 2-LTR circular DNA in peripheral blood mononuclear cells and brain tissues from SIV-infected macaques.
  
  
  
  This assay has sensitivity, accuracy and reproducibility over seven orders of magnitude. High copy numbers of SIV 2-LTR circles were correlated to high proviral DNA levels in brains of two SIV encephalitic animals. In contrast, no 2-LTR circles were detectable in two SIV-infected animals with no sign of encephalitis or two animals that had mild encephalitis with low levels of proviral DNA.
  
  
  
  These results suggest that simultaneous application of total proviral DNA and 2-LTR circle assays provides quantitative evaluation of pathogenesis and outcome of SIV infection in macaques.
authors.affiliation: Advanced BioScience Laboratories, Inc., Rockville, MD, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Lee
authors.affiliation: Department of Microbiology and Molecular Cell Biology, Eastern Virginia Medical School, Norfolk, VA, USA. kimw@evms.edu.
authors.forename: Woong-Ki
authors.initials: WK
authors.lastname: Kim
authors.affiliation: Division of Immunology, Tulane National Primate Research Center, Covington, LA, USA.
authors.forename: Marcelo J
authors.initials: MJ
authors.lastname: Kuroda
authors.affiliation: Advanced BioScience Laboratories, Inc., Rockville, MD, USA.
authors.forename: Ranajit
authors.initials: R
authors.lastname: Pal
authors.affiliation: Advanced BioScience Laboratories, Inc., Rockville, MD, USA. hye.chung@ablinc.com.
authors.forename: Hye Kyung
authors.initials: HK
authors.lastname: Chung
mesh_terms: D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000821:virology; D004270:DNA, Circular / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D007963:Leukocytes, Mononuclear / Q000821:virology; D008297:Male; D011533:Proviruses / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D016415:Sequence Alignment; D016097:Simian Acquired Immunodeficiency Syndrome / Q000821:virology*; D015302:Simian Immunodeficiency Virus / Q000235:genetics*
chemical_list: D004270:DNA, Circular; D004279:DNA, Viral
keywords: SIV infection; anti-HIV; immune activation; macaque; pathogenesis; proviral DNA; reservoir; vaccine