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1996-03-09
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Document 0078
DOCN M9650078
TI Internal controls as performance monitors and quantitative standards in
the detection by polymerase chain reaction of herpes simplex virus and
cytomegalovirus in clinical specimens.
DT 9605
AU Nash KA; Klein JS; Inderlied CB; Department of Pathology, Childrens
Hospital Los Angeles, CA; 90027, USA.
SO Mol Cell Probes. 1995 Oct;9(5):347-56. Unique Identifier : AIDSLINE
GENBANK/U06236
AB Internal controls (IC) were produced and characterized for an HSV and a
CMV PCR assay which serve both as test performance monitors and as
quantitative standards. In each of the PCR assays the IC and native
targets were amplified with equal efficiency and were detected with the
same sensitivity, i.e. < 10 target copies. An algorithm was developed
for the use of IC as a quantitative standard which entailed coamplifying
a test specimen with four two-fold dilutions of the respective IC target
(63-500 copies), followed by regression analysis of the relative yield
of amplification products. This approach allowed the determination of
both the initial virus genome copy number and the variability of the
results, which provided a confidence index for the PCR assay. The
relative yields of PCR products were determined by Southern blot and
probe hybridization and by densitometry of digitized ethidium
bromide-stained gels. Both methods produced estimations of the initial
target copy numbers within +/- 40% of the expected value. Such a
comprehensive analysis of an internal control for a PCR assay provides a
rigorous control of test performance and permits reliable quantitative
interpretation of a PCR assay result.
DE Adolescence Algorithms AIDS-Related Opportunistic Infections/DIAGNOSIS
Base Sequence Case Report Child Comparative Study
Cytomegalovirus/GENETICS/*ISOLATION & PURIF Cytomegalovirus
Infections/DIAGNOSIS DNA Primers DNA, Viral/ANALYSIS/GENETICS Herpes
Simplex/DIAGNOSIS Herpesvirus 2, Human/GENETICS/*ISOLATION & PURIF
Human Male Molecular Sequence Data Oligonucleotide Probes
*Polymerase Chain Reaction JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).