home
***
CD-ROM
|
disk
|
FTP
|
other
***
search
/
Shareware Overload Trio 2
/
Shareware Overload Trio Volume 2 (Chestnut CD-ROM).ISO
/
dir26
/
med9410l.zip
/
M94A1883.TXT
< prev
next >
Wrap
Text File
|
1994-10-24
|
3KB
|
52 lines
Document 1883
DOCN M94A1883
TI Precision and power of discrimination of a novel quantitative HIV RNA
PCR assay.
DT 9412
AU Kwok S; Christopherson C; McKinney N; Mulder J; Meyers L; Sninsky J;
Roche Molecular Systems, Alameda, CA 94501.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):43 (abstract no. 146B). Unique
Identifier : AIDSLINE ICA10/94370692
AB OBJECTIVE: Determine the precision and power of discrimination of a
simple and colorimetric HIV-1 quantitative RNA PCR assay. METHODS: A
quantitative PCR assay for HIV-1 RNA has recently been published (J.
Clin Micro., Feb 1994). To assess the variability of the assay, each
step of the assay (detection, amplification, and sample preparation) was
dissected and studied. The reproducibility of the assay was determined
by statistical analysis of duplicate standard deviations of log copies.
This data was used to calculate error rates associated with the decision
of whether two samples have different copy numbers. RESULTS: To
determine the reproducibility of the microwell detection plates,
amplified products were analyzed on duplicate wells on two plates. The
coefficient of variation (CV) of O.D.s was 2-4% within plates and no
more than 6.5% between plates. The analysis of multiple amplifications
from the same prepared samples demonstrated O.D. CVs of less than 12%,
which includes variability contributed by the detection system. To
address variability in sample preparation, quadruplicate extraction were
performed on 4 specimens. CVs of 8-30% were observed; the higher CVs
were associated with lower copy numbers (300 copies/ml). The standard
deviation of log copies (S) for the assay was determined to be 0.2
(which approximately correlates to a CV of 20%). This allows one to
recognize with 95% confidence that two samples are different if the
ratio of the copy number estimates are not between 0.57 and 1.75.
DISCUSSION AND CONCLUSION: The quantitative RNA assay described shows
good precision and can discern relatively minor changes in viremia. With
a sensitivity of 100 copies/ml, we have demonstrated that this procedure
can be used to monitor viremia in individuals a) with undetectable p24
antigen, b) with high CD4 counts, C) undergoing antiretroviral therapy,
d) post-seroconversion, and e) diagnose neonates born to infected
mothers.
DE Analysis of Variance Antiviral Agents/THERAPEUTIC USE
Colorimetry/METHODS/STATISTICS & NUMER DATA Female Human HIV Core
Protein p24/BLOOD HIV Infections/BLOOD/DRUG THERAPY/MICROBIOLOGY HIV
Seropositivity/BLOOD/DIAGNOSIS/MICROBIOLOGY HIV-1/*GENETICS/ISOLATION &
PURIF Infant, Newborn Leukocyte Count Maternal-Fetal Exchange
Polymerase Chain Reaction/*METHODS/STATISTICS & NUMER DATA Pregnancy
Pregnancy Complications, Infectious/MICROBIOLOGY RNA,
Viral/*BLOOD/*GENETICS Sensitivity and Specificity T4 Lymphocytes
Viremia/BLOOD/DRUG THERAPY/MICROBIOLOGY MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).