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. 1992 Dec;30(12):3185–3189. doi: 10.1128/jcm.30.12.3185-3189.1992

Coamplified positive control detects inhibition of polymerase chain reactions.

R W Cone 1, A C Hobson 1, M L Huang 1
PMCID: PMC270618  PMID: 1333488

Abstract

Polymerase chain reactions (PCR) may fail to react because the substrate DNA is absent (true negative) or because of inhibition of specific amplification (false negative). The use of positive controls can increase confidence in negative PCR results by ruling out failure due to inhibition as a cause of the lack of amplification products. This report describes the construction and application of coamplified positive controls for herpes simplex virus and human herpesvirus 6 amplifications. Herpes simplex virus and human herpesvirus 6 PCR products were modified to generate control PCR products in which the original probe sequences were replaced by a Drosophila probe sequence. Implementation of the coamplified controls increased our specimen throughput in comparison with the parallel control amplifications used previously. Clinical laboratories using PCR for diagnosis of infectious diseases may find positive controls particularly helpful for increasing confidence that negative amplifications represent truly negative specimens.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aurelius E., Johansson B., Sköldenberg B., Staland A., Forsgren M. Rapid diagnosis of herpes simplex encephalitis by nested polymerase chain reaction assay of cerebrospinal fluid. Lancet. 1991 Jan 26;337(8735):189–192. doi: 10.1016/0140-6736(91)92155-u. [DOI] [PubMed] [Google Scholar]
  2. Cone R. W., Hobson A. C., Huang M. L., Fairfax M. R. Polymerase chain reaction decontamination: the wipe test. Lancet. 1990 Sep 15;336(8716):686–687. doi: 10.1016/0140-6736(90)92180-p. [DOI] [PubMed] [Google Scholar]
  3. Cone R. W., Hobson A. C., Palmer J., Remington M., Corey L. Extended duration of herpes simplex virus DNA in genital lesions detected by the polymerase chain reaction. J Infect Dis. 1991 Oct;164(4):757–760. doi: 10.1093/infdis/164.4.757. [DOI] [PubMed] [Google Scholar]
  4. Dahlén P. O., Iitiä A. J., Skagius G., Frostell A., Nunn M. F., Kwiatkowski M. Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay. J Clin Microbiol. 1991 Apr;29(4):798–804. doi: 10.1128/jcm.29.4.798-804.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eggerding F. A., Peters J., Lee R. K., Inderlied C. B. Detection of rubella virus gene sequences by enzymatic amplification and direct sequencing of amplified DNA. J Clin Microbiol. 1991 May;29(5):945–952. doi: 10.1128/jcm.29.5.945-952.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gilliland G., Perrin S., Blanchard K., Bunn H. F. Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2725–2729. doi: 10.1073/pnas.87.7.2725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gopal M. R., Thomson B. J., Fox J., Tedder R. S., Honess R. W. Detection by PCR of HHV-6 and EBV DNA in blood and oropharynx of healthy adults and HIV-seropositives. Lancet. 1990 Jun 30;335(8705):1598–1599. doi: 10.1016/0140-6736(90)91433-b. [DOI] [PubMed] [Google Scholar]
  8. Israele V., Shirley P., Sixbey J. W. Excretion of the Epstein-Barr virus from the genital tract of men. J Infect Dis. 1991 Jun;163(6):1341–1343. doi: 10.1093/infdis/163.6.1341. [DOI] [PubMed] [Google Scholar]
  9. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  10. O'Hare K., Murphy C., Levis R., Rubin G. M. DNA sequence of the white locus of Drosophila melanogaster. J Mol Biol. 1984 Dec 15;180(3):437–455. doi: 10.1016/0022-2836(84)90021-4. [DOI] [PubMed] [Google Scholar]
  11. Stuve L. L., Brown-Shimer S., Pachl C., Najarian R., Dina D., Burke R. L. Structure and expression of the herpes simplex virus type 2 glycoprotein gB gene. J Virol. 1987 Feb;61(2):326–335. doi: 10.1128/jvi.61.2.326-335.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wang A. M., Doyle M. V., Mark D. F. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9717–9721. doi: 10.1073/pnas.86.24.9717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Zazzi M., Romano L., Brasini A., Valensin P. E. Low human immunodeficiency virus titer and polymerase chain reaction false-negatives. J Infect Dis. 1992 Apr;165(4):779–780. doi: 10.1093/infdis/165.4.779. [DOI] [PubMed] [Google Scholar]

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