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
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