J Med Virol. 1997 Jan;51(1):56-63. Unique Identifier : AIDSLINE
The quantification of human immunodeficiency virus type 1 (HIV-1) RNA or
hepatitis C virus (HCV) RNA has been facilitated by adapting a spin
column procedure for sample preparation and the use of chemiluminescent
detection of polymerase chain reaction PCR) products in microtiter plate
format. All materials were commercially available and relatively
inexpensive. By making a single dilution prior to amplification,
concentrations of 500 copies to 2.5 million HIV-1 1 RNA copies per mL
and 1,000 copies to 50 million HCV RNA copies per mL could be determined
on 140-microL samples. Between-run imprecision employing the improved
procedure for HIV-1 RNA was 23%. Correlation of HIV-1 RNA concentrations
obtained using chemiluminescent detection with values obtained by
colorimetric assay of PCR products was 0.98. Correlation of HCV RNA
concentration determined by the spin column-chemiluminescent assay
procedure with those obtained by branched DNA methodology was 0.91. Spin
columns could be used with serum or plasma containing
acid-citrate-dextrose or heparin anticoagulant, but heparinized samples
required treatment with heparinase prior to amplification.
*Hepatitis C/DIAGNOSIS *Hepatitis C-Like Viruses/ISOLATION & PURIF
*HIV Infections/DIAGNOSIS *HIV-1/ISOLATION & PURIF *Polymerase Chain
Reaction/METHODS *RNA, Viral/ISOLATION & PURIF *Specimen
Handling/METHODS
www.aegis.org