Monitoring of human cytomegalovirus infection in immunosuppressed patients using real-time PCR on whole blood.
BACKGROUND
Quantitative monitoring of human cytomegalovirus (HCMV) is currently used in the follow-up of immunosuppressed patients.


OBJECTIVE
To investigate whether real-time PCR quantification (QPCR) of HCMV DNA could replace pp65 antigenemia.


METHODS
We compared HCMV QPCR on whole blood (WB) and on plasma with a pp65-antigenemia assay on 192 samples. Afterwards, we tested 1310 samples from 308 immunosuppressed patients both by antigenemia assay and QPCR on WB.


RESULTS
The first study comparison showed that QPCR results on WB and plasma were significantly correlated with antigenemia. QPCR on WB was more sensitive than QPCR on plasma or antigenemia, detecting 31 and 49 additional positive samples, respectively. During the second comparison, QPCR on WB and antigenemia were again correlated (r=0.70; p<0.0001), but QPCR detected 244 additional positive samples. HCMV DNA was detected earlier than pp65 antigen (median difference: 14 days; range: 7-30). One, 5, 10, 50 and 100 pp65-positive cells/200,000 leukocytes corresponded to 439, 1531, 2623, 9150 and 15,671 HCMV DNA copies/mL of WB, respectively, but this equivalence differed according to the sub-group of patients considered.


CONCLUSIONS
QPCR on WB is the most sensitive method for the monitoring of HCMV infection in immunosuppressed patients.
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Laboratoire de Virologie du CERVI, UPRES EA 2387 Université Pierre et Marie Curie-Paris VI, Groupe Hospitalier Pitié-Salpêtrière, 75013 Paris, France. claire.deback@psl.aphp.fr
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D000328:Adult; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology; D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D016867:Immunocompromised Host; D010750:Phosphoproteins / Q000097:blood; D010949:Plasma / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000097:blood
D004279:DNA, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa