PMID- 16444011
OWN - NLM
STAT- MEDLINE
DCOM- 20060713
LR  - 20191210
IS  - 1073-6085 (Print)
IS  - 1073-6085 (Linking)
VI  - 32
IP  - 2
DP  - 2006 Feb
TI  - Fluorogenic LUX primer for quantitation of HIV-1 by real-time RT-PCR.
PG  - 101-10
AB  - Measurement of HIV-1 viral load in plasma is an important marker of disease 
      progression and efficacy of antiretroviral therapy. Real-time polymerase chain 
      reaction (PCR) offers an opportunity to develop more affordable alternative viral 
      load assays. This article reports on the development of a novel real-time 
      reverse-transcriptase (RT)-PCR assay for quantitation of HIV-1 RNA copies. This 
      assay utilizes the LightCycler (version 2) real-time PCR platform and light upon 
      extension (LUX) primer for specific detection of amplicons. An external standard 
      (ES) for quantitation of viral RNA represents an in vitro transcribed RNA. The 
      LUX assay shows a wide linear (R2 = 0.99) dynamic range from 4 x 10(6) to 4 x 
      10(2) copies/mL. Analytical sensitivity of the assay is 4 x 10(2) copies/mL of ES 
      RNA. Intra- and inter-assay variability of the LUX assay was less than 0.5log(10) 
      copies of ES RNA (i.e., no clinically significant variability was found). 
      Virology quality assurance (VQA) HIV-1 RNA copy controls were used to validate ES 
      and preliminarily evaluate the assay performance. This feasibility study 
      demonstrated that the LUX assay is sensitive, reproducible, and compares well to 
      the Roche Amplicor tests used for characterization of the RNA copy controls. 
      These results suggest further evaluation of the LUX assay using a large cohort of 
      well-characterized samples from HIV-1 positive individuals.
FAU - Rekhviashvili, Natela
AU  - Rekhviashvili N
AD  - Department of Molecular Medicine and Hematology, National Health Laboratory 
      Services, University of the Witwatersrand, Faculty of Health Science, 
      Johannesburg, South Africa. rekhn@hsc.pg.wits.ac.za
FAU - Stevens, Gwynneth
AU  - Stevens G
FAU - Scott, Lesley
AU  - Scott L
FAU - Stevens, Wendy
AU  - Stevens W
LA  - eng
GR  - 1219 AI 53217-01/AI/NIAID NIH HHS/United States
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - Switzerland
TA  - Mol Biotechnol
JT  - Molecular biotechnology
JID - 9423533
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - DNA Primers/*chemistry/*genetics
MH  - Evaluation Studies as Topic
MH  - Feasibility Studies
MH  - HIV-1/*genetics/*isolation & purification
MH  - Humans
MH  - In Vitro Techniques
MH  - RNA, Viral/blood/genetics
MH  - Reference Standards
MH  - Reproducibility of Results
MH  - Reverse Transcriptase Polymerase Chain Reaction/*instrumentation/methods
MH  - Sensitivity and Specificity
MH  - Software
MH  - Viral Load/statistics & numerical data
EDAT- 2006/01/31 09:00
MHDA- 2006/07/14 09:00
CRDT- 2006/01/31 09:00
PHST- 2006/01/31 09:00 [pubmed]
PHST- 2006/07/14 09:00 [medline]
PHST- 2006/01/31 09:00 [entrez]
AID - MB:32:2:101 [pii]
AID - 10.1385/MB:32:2:101 [doi]
PST - ppublish
SO  - Mol Biotechnol. 2006 Feb;32(2):101-10. doi: 10.1385/MB:32:2:101.