CLINICAL MICROBIOLOGY REVIEWS, July 1998, p. 533–554 Vol. 11, No. 3 0893-8512/98/$04.0010 Copyright © 1998, American Society for Microbiology. All Rights Reserved. Quantitation of Cytomegalovirus: Methodologic Aspects and Clinical Applications 1 2 MICHAEL BOECKH * AND GUY BOIVIN Fred Hutchinson Cancer Research Center, Seattle, Washington,1 and Centre Hospitalier de l’Universite´ Laval, Sainte-Foy, Quebec, Canada2 INTRODUCTION .......................................................................................................................................................534 PATHOPHYSIOLOGY OF CMV DISSEMINATION ...........................................................................................534 METHODS FOR CMV QUANTIFICATION..........................................................................................................534 Quantitative Viral Cultures...................................................................................................................................535 Principle and assay characteristics..................................................................................................................535 Assay performance..............................................................................................................................................535 Reproducibility and test accuracy ....................................................................................................................535 Summary ..............................................................................................................................................................535 Quantitative pp65 Antigenemia Assay .................................................................................................................536 Principle ...............................................................................................................................................................536 Assay characteristics ..........................................................................................................................................536 (i) Isolation and preparation of slides ........................................................................................................536 (ii) Fixation......................................................................................................................................................536 (iii) Immunostaining ......................................................................................................................................536 (iv) Quantification ..........................................................................................................................................538 Assay performance..............................................................................................................................................538 Reproducibility and test accuracy ....................................................................................................................538 Summary ..............................................................................................................................................................538 Quantitative PCR Assays.......................................................................................................................................538 General principles ..............................................................................................................................................538 Protocols and assay performance.....................................................................................................................540 Reproducibility and test accuracy ....................................................................................................................541 Summary ..............................................................................................................................................................541 Branched-DNA Signal Amplification Assay ........................................................................................................541 Principle ...............................................................................................................................................................541 Assay characteristics ..........................................................................................................................................541 Assay performance..............................................................................................................................................541 Reproducibility and test accuracy ....................................................................................................................541 Summary ..............................................................................................................................................................541 Hybrid Capture CMV DNA Assay........................................................................................................................541 Principle ...............................................................................................................................................................541 Assay characteristics ..........................................................................................................................................541 Assay performance..............................................................................................................................................542 Reproducibility and assay accuracy .................................................................................................................542 Summary ..............................................................................................................................................................542 CORRELATION OF SYSTEMIC VIRAL LOAD AND CMV DISEASE ............................................................542 Solid-Organ Transplantation................................................................................................................................542 Relationship between systemic viral load and CMV disease........................................................................542 Summary ..............................................................................................................................................................542 Marrow and Stem Cell Transplantation .............................................................................................................542 Role of viremia ....................................................................................................................................................542 Relationship of systemic viral load and CMV disease ..................................................................................543 Early antiviral treatment based on viral load ................................................................................................544 Systemic viral load in autologous transplantation ........................................................................................545 Summary ..............................................................................................................................................................545 HIV and AIDS.........................................................................................................................................................545 Relationship of systemic viral load and CMV disease ..................................................................................545 Summary ..............................................................................................................................................................546 * Corresponding author. Mailing address: Program in Infectious Diseases, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. North, Seattle, WA 98109. Phone: (206) 667-6702. Fax: (206) 667-4411. E-mail: [email protected]. 533 534 BOECKH AND BOIVIN CLIN.MICROBIOL.REV. RESPONSE TO TREATMENT, SURROGATE MARKER FOR ANTIVIRAL RESISTANCE .......................546 Solid-Organ Transplantation................................................................................................................................546 Marrow and Stem Cell Transplantation .............................................................................................................547 HIV and AIDS.........................................................................................................................................................547 Systemic Viral Load and Resistance....................................................................................................................547 Relationship between systemic viral load and resistance .............................................................................547 Summary ..............................................................................................................................................................548 VIRAL LOAD AT SPECIFIC ORGAN SITES .......................................................................................................548 Central Nervous System.........................................................................................................................................548 Lungs