title: Quantification of HIV-1 RNA load by one-tube-one-step RT PCR and real time PCR assay with TaqMan probe.
abstract: OBJECTIVE
  To develop a less expensive assay to calculate HIV-1 viral load for use in resource-limited countries.
  
  
  METHODS
  An In-house One-tube-one-step Viral Load Assay (IOVA) was developed by using real-time PCR-based with TaqMan probe. Primers and probe were designed from the conserved region of sequences from all HIV subtypes. A standard curve was generated from reference virus in various dilutions. IOVA was applied on 105 HIV-positive and 25 HIV-negative samples and compared with the results from ROCHE AMPLICLOR.
  
  
  RESULTS
  IOVA measured HIV RNA in the samples ranging from 125 to 2 x 10(6) copies/mL. The coefficient of variation of intra- and inter-assay ranged from 0.68% to 7.89%. The sensitivity, specificity, positive and negative predictive values were 92%, 100%, 100% and 79.5% respectively. The parallel quantitative analysis showed high correlation (r=0.95) between IOVA and AMPLICOR.
  
  
  CONCLUSIONS
  A new HIV-1 viral load assay was developed and validated. It was reliable and less expensive.
authors.affiliation: Deparment of Pediatrics, Phramongkutklao Hospital, Bangkok, Thailand. veerachaiw@yahoo.com
authors.forename: Veerachai
authors.initials: V
authors.lastname: Watanaveeradej
authors.forename: Sudalak
authors.initials: S
authors.lastname: Sirirattanaphoomee
authors.forename: Wasun
authors.initials: W
authors.lastname: Chantratita
authors.forename: Sorachai
authors.initials: S
authors.lastname: Nitayaphan
authors.forename: Kwanjai
authors.initials: K
authors.lastname: Viputtikul
authors.forename: Angkool
authors.initials: A
authors.lastname: Kerdpanich
authors.forename: Rudiwilai
authors.initials: R
authors.lastname: Samakoses
authors.forename: Sriluk
authors.initials: S
authors.lastname: Simasathien
mesh_terms: D000328:Adult; D002648:Child; D015658:HIV Infections / Q000097:blood*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D015347:RNA Probes; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000379:methods*
chemical_list: D015347:RNA Probes; D012367:RNA, Viral