| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
University Division of Communicable Diseases, Royal Free Hospital School of Medicine, Rowland Hill Street, London NW3 2PF, U.K.
The important goal of developing quantitative assays for viral nucleic acids in clinical samples has been achieved for human cytomegalovirus (HCMV) by using a modified polymerase chain reaction (PCR). A control PCR target sequence was constructed by PCR mutagenesis to allow the post-amplification quantification of HCMV DNA. The control region was identical to a naturally occurring sequence within the glycoprotein B (gB) coding part of the virus genome, except that a unique restriction site, introduced by the aforementioned mutagenesis step, allowed post-amplification differentiation of control/non-control target amplified product. This technique was initially validated using known amounts of cloned control/non-control target DNA, and was found to be sufficiently sensitive to allow the quantification of a range of 10 to 106 genome equivalents of virus. The method was applied to urine samples of congenitally infected infants for which infectious virus titres were available. The results obtained demonstrated that the number of infectious virions determined by conventional cell culture represented a small proportion of the HCMV genome present in the samples, as assessed by the quantitative PCR methodology.
Received 8 October 1991;
accepted 8 May 1992.
This article has been cited by other articles:
|
|
 |
|
  S Walter, C Atkinson, M Sharland, P Rice, E Raglan, V C Emery, and P D Griffiths Congenital cytomegalovirus: association between dried blood spot viral load and hearing loss Arch. Dis. Child. Fetal Neonatal Ed., July 1, 2008; 93(4): F280 - F285. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Herrmann, V. Cavaglia Larsson, C.-J. Rubin, F. Sund, B.-M. Eriksson, J. Arvidson, Z. Yun, K. Bondeson, and J. Blomberg Comparison of a Duplex Quantitative Real-Time PCR Assay and the COBAS Amplicor CMV Monitor Test for Detection of Cytomegalovirus J. Clin. Microbiol., May 1, 2004; 42(5): 1909 - 1914. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Weinberg, D. Schissel, and R. Giller Molecular Methods for Cytomegalovirus Surveillance in Bone Marrow Transplant Recipients J. Clin. Microbiol., November 1, 2002; 40(11): 4203 - 4206. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. G. Revello and G. Gerna Diagnosis and Management of Human Cytomegalovirus Infection in the Mother, Fetus, and Newborn Infant Clin. Microbiol. Rev., October 1, 2002; 15(4): 680 - 715. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. F. L. Fryer, P. D. Griffiths, J. A. Fishman, V. C. Emery, and D. A. Clark Quantitation of Porcine Cytomegalovirus in Pig Tissues by PCR J. Clin. Microbiol., March 1, 2001; 39(3): 1155 - 1156. [Abstract] [Full Text]
|
|
|
|
 |
|
 F M Mattes, J E McLaughlin, V C Emery, D A Clark, and P D Griffiths Histopathological detection of owl's eye inclusions is still specific for cytomegalovirus in the era of human herpesviruses 6 and 7 J. Clin. Pathol., August 1, 2000; 53(8): 612 - 614. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Gerna, F. Baldanti, D. Lilleri, M. Parea, E. Alessandrino, A. Pagani, F. Locatelli, J. Middeldorp, and M. G. Revello Human Cytomegalovirus Immediate-Early mRNA Detection by Nucleic Acid Sequence-Based Amplification as a New Parameter for Preemptive Therapy in Bone Marrow Transplant Recipients J. Clin. Microbiol., May 1, 2000; 38(5): 1845 - 1853. [Abstract] [Full Text]
|
|
|
|
|
|
A. Weinberg, T. N. Hodges, S. Li, G. Cai, and M. R. Zamora Comparison of PCR, Antigenemia Assay, and Rapid Blood Culture for Detection and Prevention of Cytomegalovirus Disease after Lung Transplantation J. Clin. Microbiol., February 1, 2000; 38(2): 768 - 772. [Abstract] [Full Text]
|
|
|
|
 |
|
 V. C. Emery, A. V. Cope, E. F. Bowen, D. Gor, and P. D. Griffiths The Dynamics of Human Cytomegalovirus Replication In Vivo J. Exp. Med., July 19, 1999; 190(2): 177 - 182. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Gerna, F. Baldanti, J. M. Middeldorp, M. Furione, M. Zavattoni, D. Lilleri, and M. G. Revello Clinical Significance of Expression of Human Cytomegalovirus pp67 Late Transcript in Heart, Lung, and Bone Marrow Transplant Recipients as Determined by Nucleic Acid Sequence-Based Amplification J. Clin. Microbiol., April 1, 1999; 37(4): 902 - 911. [Abstract] [Full Text]
|
|
|
|
|
|
R. B. Domingues, F. D. Lakeman, M. S. Mayo, and R. J. Whitley Application of Competitive PCR to Cerebrospinal Fluid Samples from Patients with Herpes Simplex Encephalitis J. Clin. Microbiol., August 1, 1998; 36(8): 2229 - 2234. [Abstract] [Full Text]
|
|
|
|
|
|
M. Boeckh and G. Boivin Quantitation of Cytomegalovirus: Methodologic Aspects and Clinical Applications Clin. Microbiol. Rev., July 1, 1998; 11(3): 533 - 554. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A. Fishman and R. H. Rubin Infection in Organ-Transplant Recipients N. Engl. J. Med., June 11, 1998; 338(24): 1741 - 1751. [Full Text] [PDF]
|
|
|
|
|
|
G. Gerna, M. Zavattoni, E. Percivalle, P. Grossi, M. Torsellini, and M. G. Revello Rising Levels of Human Cytomegalovirus (HCMV) Antigenemia during Initial Antiviral Treatment of Solid-Organ Transplant Recipients with Primary HCMV Infection J. Clin. Microbiol., April 1, 1998; 36(4): 1113 - 1116. [Abstract] [Full Text]
|
|
|
|
 |
|
 N. Noraz, J. L. Lathey, and S. A. Spector Human Cytomegalovirus-Associated Immunosuppression Is Mediated Through Interferon-alpha Blood, April 1, 1997; 89(7): 2443 - 2452. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
