title: Establishment of cheap and reliable real-time PCR for quantitation of HIV-1 viral load in plasma.
abstract: OBJECTIVE
  To establish an inexpensive and reliable real-time PCR for quantitation of HIV-1 RNA from plasma samples.
  
  
  METHODS
  Previously analyzed 145 HIV-1 positive plasma samples with viral load ranging from less than 40 to approximately 1,000,000 copies/ml were included in the present study. HIV-1 gag gene was amplified and cloned into TA cloning vector External standard curve was plotted using in vitro transcribed HIV-1 RNA and utilizedfor viral quantitation in the samples. Scramble nucleotides located in HIV-1 specific probe was subsequently constructed and used for individual systemic control. The correlation coefficient and Bland-Altman plot were applied for statistical analysis of the two methods.
  
  
  RESULTS
  The limit of quantitation of the validated assay was 31 copies/ml and the linear range was approximate 31-1 x 10(7) copies/ml. After reproducibility determination using intra- and inter-run assay, it was implied that the coefficient of variation (%CV) was significantly increased while the low copy number of RNA was examined A highly correlation (r2 = 0.8099) and good agreement were obtained when the two assays were compared
  
  
  CONCLUSIONS
  Developed real-time PCR was inexpensive and reliable for quantitation of HIV-1 viral load in plasma.
authors: Division of Clinical Microbiology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
authors: Kanittaporn
authors: K
authors: Supadej
authors: Sorasak
authors: S
authors: Intorasoot
mesh_terms: D015658:HIV Infections / Q000097:blood; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D012367:RNA, Viral / Q000097:blood*; D060888:Real-Time Polymerase Chain Reaction; D019562:Viral Load; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics
chemical_list: D012367:RNA, Viral; D054301:gag Gene Products, Human Immunodeficiency Virus