Evaluation of the AmpliSensor PCR and the SHARP signal detection system for the early prediction of symptomatic CMV infection in solid transplant recipients

• Published in: Journal of clinical virology Vol. 13; no. 1; pp. 81 - 94
• Main Authors: Rautenberg, Peter, Lübbert, Christoph, Weers, Wiebke, Boetel, Eric, Schweichler, Jörg, Zhou, Lin, Costard-Jäckle, Angelika, Kraemer-Hansen, Heike, Harder, Timm C
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 01-06-1999; Elsevier Science
• Subjects: Adult; AmpliSensor PCR; Antibodies, Viral - blood; Antibodies, Viral - immunology; Biological and medical sciences; Cohort Studies; Cytomegalovirus - genetics; Cytomegalovirus - immunology; Cytomegalovirus - isolation & purification; Cytomegalovirus Infections - diagnosis; Cytomegalovirus Infections - drug therapy; Cytomegalovirus Infections - immunology; Cytomegalovirus Infections - virology; Detection system; Evaluation Studies as Topic; Human viral diseases; Humans; Immediate-Early Proteins - genetics; Infectious diseases; Medical sciences; Membrane Glycoproteins; Organ Transplantation; Phosphoproteins - immunology; Polymerase Chain Reaction - methods; Postoperative Complications - virology; Prospective Studies; Quantitative PCR monitoring; Reagent Kits, Diagnostic; Sensitivity and Specificity; SHARP signal; Solid organ recipients; Trans-Activators; Viral diseases; Viral diseases of the lymphoid tissue and the blood. Aids; Viral Envelope Proteins; Viral Matrix Proteins - immunology; Viral Proteins; Quantitative PCR monitoring; SHARP signal; Solid organ recipients; AmpliSensor PCR; Detection system; Performance evaluation; Human; Herpesviridae; Recipient; Transplantation; Method; Betaherpesvirinae; Human cytomegalovirus; Infection; Virus; Polymerase chain reaction; DNA; Viral disease; Complication; Diagnosis; Detection; Viral load; Quantitative analysis; Organ
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/S1386-6532(99)00013-X

Background: Cytomegalovirus (CMV) is associated with high morbidity and mortality in transplant patients. Specific antiviral treatment at an early stage of CMV infection may effectively ameliorate, but not eliminate CMV disease in these patients. Presently, the pp65 antigenemia test on peripheral leukocytes is the method most widely used for predicting and monitoring transplant patients for active CMV infection. Nucleic acid amplification methods are less well defined since they lack standardisation.
Objective: A seminested fluorometric PCR assay (AmpliSensor-CMV, BAG, Germany) and a one-step PCR with a signal-amplification step (SHARP, Abbott, Germany) specific for the fragments of the CMV UL 122 and UL 123 genes, respectively, were evaluated for the early diagnosis of CMV infection.
Design: A total of 26 recipients of heterogeneous solid organs were monitored prospectively for a median of 99 days after transplantation. By testing 371 clinical samples parallel with the pp65-antigen assay and IgM and IgG EIA assays the sensitivity, specificity, correlation and quantitation potential of both PCRs was evaluated.
Results: Eight out of 26 patients developed active CMV infection. A total of 48 samples of these patients exceeded a CMV-DNA load threshold of 15 genome equivalents/10
5 leukocytes (AmpliSensor-CMV) and 41 samples exceeded the critical cut-off for the SHARP system. The AmpliSensor PCR exceeded its threshold consistently before the clinical onset of CMV disease (median 8 days). There was very good agreement between symptomatic CMV infection in patients and AmpliSensor-PCR, SHARP PCR, and pp65-antigen results (
κ-coefficient>0.900). IgM and IgG EIA showed moderate agreement (
κ-coefficient=0.591 and 0.552, respectively).
Conclusion: Both PCRs and pp65 antigen assay correlated significantly better with CMV disease than serodiagnosis. The AmpliSensor PCR allowed more precisely than the SHARP system a quantitative determination of viral load and an early and reliable prediction of active CMV infection. The use of AmpliSensor PCR may improve the diagnosis and management of active CMV infection in organ transplant recipients.