Background: EBVrelated posttransplant lymphoproliferative diseases are usually accompanied by increased EBV DNA in peripheral blood. Monitoring EBV DNAemia is the basis for weighing decisions regarding initiation of preemptive or antiEBVrelated tumor therapy. However, the definition of clinically relevant cutoff values is hampered by the lack of standardization in EBV DNA testing. Objectives: To estimate interlaboratory variability and to evaluate the impact of different matrices in EBV DNA load determination in Italian laboratories involved in monitoring of virus infections in transplanted patients. Study design: Two different proficiency panels were distributed among seven centers: the first contained cellassociated and cellfree EBVs; the second was prepared by spiking both cellassociated and cellfree EBVs in EBV DNAnegative whole blood from EBV seropositive healthy donors. Samples were extracted and amplified with different methods. Intralaboratory and interlaboratory variabilities was evaluated. Results: 337 EBV DNA determinations were performed. Sensitivity was 100% for both panels, specificity was 100% for the first and 74% for the second panel, where whole blood was utilized as the matrix. Discrepant results in the second panel were restricted to samples containing low copy numbers. Quantification fell within ±0.5 log in 73% of the determinations. Values for cellassociated samples tended to be more heterogeneous than those obtained from cellfree samples. Good overall linearity was observed for each sample type; interlaboratory variability ranged from 4.71% to 12.86%. Conclusions: The results of this multicenter study indicate that EBV DNAemia may be reliably quantified by different laboratories using a variety of commercial and inhouse molecular assays.
Multicenter comparative study of Epstein-Barr virus DNA quantification for virological monitoring in transplanted patients / Abbate I.; Zanchetta M.; Gatti M.; Gabrielli L.; Milia M.G.; Lazzarotto T.; Tedeschi R.; Ghisetti V.; Clementi M.; De Rossi A.; Capobianchi M.R.. - In: JOURNAL OF CLINICAL VIROLOGY. - ISSN 1386-6532. - STAMPA. - 50:3(2011), pp. 224-229. [10.1016/j.jcv.2010.12.002]
Multicenter comparative study of Epstein-Barr virus DNA quantification for virological monitoring in transplanted patients.
LAZZAROTTO, TIZIANA;Tedeschi R.;
2011
Abstract
Background: EBVrelated posttransplant lymphoproliferative diseases are usually accompanied by increased EBV DNA in peripheral blood. Monitoring EBV DNAemia is the basis for weighing decisions regarding initiation of preemptive or antiEBVrelated tumor therapy. However, the definition of clinically relevant cutoff values is hampered by the lack of standardization in EBV DNA testing. Objectives: To estimate interlaboratory variability and to evaluate the impact of different matrices in EBV DNA load determination in Italian laboratories involved in monitoring of virus infections in transplanted patients. Study design: Two different proficiency panels were distributed among seven centers: the first contained cellassociated and cellfree EBVs; the second was prepared by spiking both cellassociated and cellfree EBVs in EBV DNAnegative whole blood from EBV seropositive healthy donors. Samples were extracted and amplified with different methods. Intralaboratory and interlaboratory variabilities was evaluated. Results: 337 EBV DNA determinations were performed. Sensitivity was 100% for both panels, specificity was 100% for the first and 74% for the second panel, where whole blood was utilized as the matrix. Discrepant results in the second panel were restricted to samples containing low copy numbers. Quantification fell within ±0.5 log in 73% of the determinations. Values for cellassociated samples tended to be more heterogeneous than those obtained from cellfree samples. Good overall linearity was observed for each sample type; interlaboratory variability ranged from 4.71% to 12.86%. Conclusions: The results of this multicenter study indicate that EBV DNAemia may be reliably quantified by different laboratories using a variety of commercial and inhouse molecular assays.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
