Substitution of a commercially available, RNA extraction procedure in an HIV-1 genotyping system improves sensitivity and allows reduced sample volume.
The use is described of a commercially available, silica-based extraction procedure for HIV-1 RNA that can be substituted for the extraction procedure supplied with a commercially available, PCR-based genotyping kit for HIV-1. The advantages of using this alternative, commercially available extraction procedure include the following: (1) reduced safety concerns, due to inactivation of virus in the initial step of the extraction procedure; (2) enhanced sensitivity, allowing the use of plasma with HIV-1 RNA levels of less than 2000 copies/ml; (3) improved yield, allowing a 60% reduction in plasma volume; and (4) convenience and improved reproducibility, with a single extraction providing RNA suitable for PCR-based sequencing and for quantitation of HIV-1 RNA levels.
Ross
L
L
Department of Virology, Glaxo Wellcome Incorporated, 5 Moore Drive, Research Triangle Park, NC 27709-3398, USA. llr8156@glaxowellcome.com
Vavro
C L
CL
Kehne
S L
SL
McClernon
D R
DR
St  Clair
M
M
D005838:Genotype; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000145:classification* / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic