This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Schaade, L.
Right arrow Articles by Kleines, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Schaade, L.
Right arrow Articles by Kleines, M.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, November 2000, p. 4006-4009, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Detection of Cytomegalovirus DNA in Human Specimens by LightCycler PCR

Lars Schaade, Peter Kockelkorn, Klaus Ritter, and Michael Kleines*

Division of Virology, Department of Medical Microbiology, University Hospital, RWTH Aachen, D-52057 Aachen, Germany

Received 10 April 2000/Returned for modification 26 June 2000/Accepted 14 August 2000

Detection of human cytomegalovirus (CMV) DNA in clinical specimens is considered a cornerstone in the diagnosis of CMV disease. The aim of this study was to evaluate a newly designed LightCycler-based quantitative CMV PCR. Specimens of human origin (n = 200) were tested using the LightCycler PCR, the quantitative COBAS AMPLICOR CMV MONITOR (CACM) assay, and a qualitative in-house PCR assay for the presence of CMV DNA. Samples that were reactive in at least two of the three assays were considered CMV DNA positive (n = 95 [47.5%]), while samples that were nonreactive in two of the three assays were considered CMV DNA negative (n = 105 [52.5%]). Using the LightCycler assay, CMV DNA was detected in 91 of the 95 CMV DNA-positive human specimens (sensitivity, 95.8%; 95% confidence interval [CI], 89.6 to 98.8) and in 1 of the CMV DNA-negative specimens (specificity, 99%; 95% CI, 94.8 to 99.8). Results of CMV load determination as assessed by both quantitative test systems were correlated (r = 0.73; P < 0.0001; 95% CI, 0.61 to 0.81). Results for undiluted samples containing a high CMV load were more accurate with the LightCycler test than were results obtained with the CACM test, which underestimated the viral load of samples containing high DNA copy numbers. The high level of sensitivity, specificity, accuracy, and rapidity provided by the LightCycler technology are favorable for the use of this system in the detection of CMV DNA in clinical specimens.

* Corresponding author. Mailing address: Division of Virology, Department of Medical Microbiology, University Hospital, RWTH Aachen, D-52057 Aachen, Germany. Phone: 49 241 80 88460. Fax: 49 241 88 88483. E-mail: mkleines{at}post.klinikum.rwth-aachen.de.

Journal of Clinical Microbiology, November 2000, p. 4006-4009, Vol. 38, No. 11
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Pang, X., Humar, A., Preiksaitis, J. K. (2008). Concurrent Genotyping and Quantitation of Cytomegalovirus gB Genotypes in Solid-Organ-Transplant Recipients by Use of a Real-Time PCR Assay. J. Clin. Microbiol. 46: 4004-4010 [Abstract] [Full Text]  
  • Gimeno, C., Solano, C., Latorre, J. C., Hernandez-Boluda, J. C., Clari, M. A., Remigia, M. J., Furio, S., Calabuig, M., Tormo, N., Navarro, D. (2008). Quantification of DNA in Plasma by an Automated Real-Time PCR Assay (Cytomegalovirus PCR Kit) for Surveillance of Active Cytomegalovirus Infection and Guidance of Preemptive Therapy for Allogeneic Hematopoietic Stem Cell Transplant Recipients. J. Clin. Microbiol. 46: 3311-3318 [Abstract] [Full Text]  
  • Garrigue, I., Doussau, A., Asselineau, J., Bricout, H., Couzi, L., Rio, C., Merville, P., Fleury, H., Lafon, M.-E., Thiebaut, R. (2008). Prediction of Cytomegalovirus (CMV) Plasma Load from Evaluation of CMV Whole-Blood Load in Samples from Renal Transplant Recipients. J. Clin. Microbiol. 46: 493-498 [Abstract] [Full Text]  
  • Hymas, W. C., Aldous, W. K., Taggart, E. W., Stevenson, J. B., Hillyard, D. R. (2008). Description and Validation of a Novel Real-Time RT-PCR Enterovirus Assay. Clin. Chem. 54: 406-413 [Abstract] [Full Text]  
  • Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
  • Nye, M. B., Leman, A. R., Meyer, M. E., Menegus, M. A., Rothberg, P. G. (2005). Sequence Diversity in the Glycoprotein B Gene Complicates Real-Time PCR Assays for Detection and Quantification of Cytomegalovirus. J. Clin. Microbiol. 43: 4968-4971 [Abstract] [Full Text]  
  • Habbal, W., Monem, F., Gartner, B. C. (2005). Errors in Published Sequences of Human Cytomegalovirus Primers and Probes: Do We Need More Quality Control?. J. Clin. Microbiol. 43: 5408-5409 [Full Text]  
  • Tang, Y.-W., Sefers, S. E., Li, H., Kohn, D. J., Procop, G. W. (2005). Comparative Evaluation of Three Commercial Systems for Nucleic Acid Extraction from Urine Specimens. J. Clin. Microbiol. 43: 4830-4833 [Abstract] [Full Text]  
  • Herrmann, B., Cavaglia Larsson, V., Rubin, C.-J., Sund, F., Eriksson, B.-M., Arvidson, J., Yun, Z., Bondeson, K., Blomberg, J. (2004). Comparison of a Duplex Quantitative Real-Time PCR Assay and the COBAS Amplicor CMV Monitor Test for Detection of Cytomegalovirus. J. Clin. Microbiol. 42: 1909-1914 [Abstract] [Full Text]  
  • Hong, K. M., Najjar, H., Hawley, M., Press, R. D. (2004). Quantitative Real-Time PCR with Automated Sample Preparation for Diagnosis and Monitoring of Cytomegalovirus Infection in Bone Marrow Transplant Patients. Clin. Chem. 50: 846-856 [Abstract] [Full Text]  
  • Kleines, M., Schellenberg, K., Ritter, K. (2003). Efficient Extraction of Viral DNA and Viral RNA by the Chemagic Viral DNA/RNA Kit Allows Sensitive Detection of Cytomegalovirus, Hepatitis B Virus, and Hepatitis G Virus by PCR. J. Clin. Microbiol. 41: 5273-5276 [Abstract] [Full Text]  
  • Pang, X. L., Chui, L., Fenton, J., LeBlanc, B., Preiksaitis, J. K. (2003). Comparison of LightCycler-Based PCR, COBAS Amplicor CMV Monitor, and pp65 Antigenemia Assays for Quantitative Measurement of Cytomegalovirus Viral Load in Peripheral Blood Specimens from Patients after Solid Organ Transplantation. J. Clin. Microbiol. 41: 3167-3174 [Abstract] [Full Text]  
  • Leruez-Ville, M., Ouachee, M., Delarue, R., Sauget, A.-S., Blanche, S., Buzyn, A., Rouzioux, C. (2003). Monitoring Cytomegalovirus Infection in Adult and Pediatric Bone Marrow Transplant Recipients by a Real-Time PCR Assay Performed with Blood Plasma. J. Clin. Microbiol. 41: 2040-2046 [Abstract] [Full Text]  
  • Li, H., Dummer, J. S., Estes, W. R., Meng, S., Wright, P. F., Tang, Y.-W. (2003). Measurement of Human Cytomegalovirus Loads by Quantitative Real-Time PCR for Monitoring Clinical Intervention in Transplant Recipients. J. Clin. Microbiol. 41: 187-191 [Abstract] [Full Text]  
  • Stocher, M., Berg, J. (2002). Normalized Quantification of Human Cytomegalovirus DNA by Competitive Real-Time PCR on the LightCycler Instrument. J. Clin. Microbiol. 40: 4547-4553 [Abstract] [Full Text]  
  • Miller, N., Cleary, T., Kraus, G., Young, A. K., Spruill, G., Hnatyszyn, H. J. (2002). Rapid and Specific Detection of Mycobacterium tuberculosis from Acid-Fast Bacillus Smear-Positive Respiratory Specimens and BacT/ALERT MP Culture Bottles by Using Fluorogenic Probes and Real-Time PCR. J. Clin. Microbiol. 40: 4143-4147 [Abstract] [Full Text]  
  • Piiparinen, H., Hockerstedt, K., Lappalainen, M., Suni, J., Lautenschlager, I. (2002). Monitoring of Viral Load by Quantitative Plasma PCR during Active Cytomegalovirus Infection of Individual Liver Transplant Patients. J. Clin. Microbiol. 40: 2945-2952 [Abstract] [Full Text]  
  • Mackay, I. M., Arden, K. E., Nitsche, A. (2002). Real-time PCR in virology. Nucleic Acids Res 30: 1292-1305 [Abstract] [Full Text]  
  • Whiley, D. M., Mackay, I. M., Sloots, T. P. (2001). Detection and Differentiation of Human Polyomaviruses JC and BK by LightCycler PCR. J. Clin. Microbiol. 39: 4357-4361 [Abstract] [Full Text]  
  • Schaade, L., Kockelkorn, P., Ritter, K., Kleines, M. (2001). Detection of Cytomegalovirus DNA in Human Specimens by LightCycler PCR: Melting Point Analysis Is Mandatory To Detect Virus Strains with Point Mutations in the Target Sequence of the Hybridization Probes. J. Clin. Microbiol. 39: 3809-3809 [Full Text]  
  • Read, S. J., Mitchell, J. L., Fink, C. G. (2001). LightCycler Multiplex PCR for the Laboratory Diagnosis of Common Viral Infections of the Central Nervous System. J. Clin. Microbiol. 39: 3056-3059 [Abstract] [Full Text]  
  • Kearns, A. M., Draper, B., Wipat, W., Turner, A. J. L., Wheeler, J., Freeman, R., Harwood, J., Gould, F. K., Dark, J. H., Schaade, L., Kleines, M. (2001). LightCycler-Based Quantitative PCR for Detection of Cytomegalovirus in Blood, Urine, and Respiratory Samples. J. Clin. Microbiol. 39: 2364-2365 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»