Evaluation of real-time PCR assay for early detection of Cytomegalovirus infection in solid-organ transplant recipients
Abstract number: P996
Pape M., Mandraveli K., Papadopoulou D., Kazakos E.I., Alexiou-Daniel S.
Objective: The aim of this study was to evaluate the advantages of Real-time PCR for early detection of CMV infection in solid-organ transplant recipients by comparison to four serology methods such as ELISA, Capture-ELISA, IFA, Western-blot.
Methods: The study included 90 solid-organ transplant recipients with clinical manifestations of recent CMV infection. Whole blood specimens and sera were collected from all patients. The whole blood specimens were processed with Real-time PCR (artus CMV LC PCR, QIAGEN) to detect CMV viral load, while the same assay was employed to detect EBV (LC EBV Quantification, Roche) and HSV/VZV (artus LC, QIAGEN) in 67 and 38 samples, respectively. All sera were tested for anti-Cytomegalovirus IgG/IgM antibodies by ELISA (AXSYM Abbott). IgM positive results were confirmed by Capture-ELISA (CE-Bouty), indirect immunofluorescene (IFA, Focus) and Western-blot (Wb-Immunogenetics) assays.
Results: CMV viral load was detected in 20 patients, while 7 of these patients had evidence of EBV co-infection(PCR+). EBV viral load was detected in 14 specimens, but no HSV or VZV DNA was detected. ELISA revealed an anti-CMV IgG positive rate of 90% in both PCR positive and negative patients. IgM antibodies were present in 15/20 (75%) CMV PCR positive patients and in 14/70 (20%) CMV PCR negative patients. The positive rates of the rest serology assays for the PCR(+) and ELISA IgM(+) patients were Capture Elisa (+) 84%, IFA (+) 73%, Western-blot (+) 88%, while the respective rates for the PCR(-) and ELISA IgM(+) patients were Capture Elisa (+) 50%, IFA (+) 35%, Western-blot (+) 43%. High viral load (>103 copies/ml) was detected in 10/15 patients with ELISA IgM positive results. Low viral load (<103 copies/ml) was detected in 5/20 patients with ELISA IgM negative results
Conclusion: The referring doctors are requested to provide more clinical informations. Double samples should be tested by all serology assays in order to access a safe diagnosis and eliminate the possibility of false negative and positive results. Evaluation of viral load by Real-time PCR seems to be more sensitive and specific for the early detection of CMV infection than the serology assays.
|Session name:||18th European Congress of Clinical Microbiology and Infectious Diseases|
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