Evaluation of the AmpliSensor PCR and the SHARP signal detection system for the early prediction of symptomatic CMV infection in solid transplant recipients
P. Rautenberg et al., Evaluation of the AmpliSensor PCR and the SHARP signal detection system for the early prediction of symptomatic CMV infection in solid transplant recipients, J CLIN VIRO, 13(1-2), 1999, pp. 81-94
Background: Cytomegalovirus (CMV) is associated with high morbidity and mor
tality in transplant patients. Specific antiviral treatment at an early sta
ge of CMV infection may effectively ameliorate, but not eliminate CMV disea
se in these patients. Presently, the pp65 antigenemia test on peripheral le
ukocytes is the method most widely used for predicting and monitoring trans
plant 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, Germa
ny) and a one-step PCR with a signal-amplification step (SHARP, Abbott, Ger
many) specific for the fragments of the CMV UL 122 and UL 123 genes, respec
tively, were evaluated for the early diagnosis of CMV infection.
Design: A total of 26 recipients of heterogeneous solid organs were monitor
ed prospectively for a median of 99 days after transplantation. By testing
371 clinical samples parallel with the pp65-antigen assay and IgM and IgG E
IA assays the sensitivity, specificity, correlation and quantitation potent
ial of both PCRs was evaluated.
Results: Eight out of 26 patients developed active CMV infection. A total o
f 48 samples of these patients exceeded a CMV-DNA load threshold of 15 geno
me equivalents/10(5) leukocytes (AmpliSensor-CMV) and 41 samples exceeded t
he critical cut-off for the SHARP system. The AmpliSensor PCR exceeded its
threshold consistently before the clinical onset of CMV disease (median 8 d
ays). There was very good agreement between symptomatic CMV infection in pa
tients and AmpliSensor-PCR, SHARP PCR, and pp65-antigen results (k-coeffici
ent > 0.900). IgM and Ige EIA showed moderate agreement (k-coefficient = 0.
591 and 0.552, respectively).
Conclusion: Both PCRs and pp65 antigen assay correlated significantly bette
r with CMV disease than serodiagnosis. The AmpliSensor PCR allowed more pre
cisely than the SHARP system a quantitative determination of viral load and
an early and reliable prediction of active CMV infection. The use of Ampli
Sensor PCR may improve the diagnosis and management of active CMV infection
in organ transplant recipients. (C) 1999 Elsevier Science B.V. All rights
reserved.