Data di Pubblicazione:
2010
Citazione:
Comparison of three real-time pcrs for the quantification of CMV-DNA on dbs / L. Bubba, A. Mammoliti, S. Binda, M. Gambino, L. Pellegrinelli, V. Primache, M. Barbi. ((Intervento presentato al 3. convegno Congenital Cytomegalovirus Conference tenutosi a Paris nel 2010.
Abstract:
BRIEF INTRODUCTION
The Dried Blood Spot (DBS) is a useful sample for diagnosing congenital CMV infection (cCMV). Some
authors suggest that a high viral load in blood samples could have an unfavourable prognostic value.
In this study we compared the performances of three methods in measuring CMV viral load in DBS.
MATERIALS AND METHODS
We tested 36 DBS samples from QCMD panels (2006-2009). Purification of DNA was performed with a
commercial kit (QIAGEN), following the DBS protocol. Viral DNA was quantified by means of an
in-house method (gB gene) and two commercial kits “CMV, HHV6, 7, 8 R-gene”, Argene (for CMV:
ppUL83) and “Q-CMV Real-time Complete kit”, Nanogen (MIEA).
CLINICAL CASES OR SUMMARY RESULTS
Intra- and inter- assay viral load values were within ±0.5log10 unit. All DBS prepared with whole
blood spiked with 1E+02 copies/ml of CMV-DNA tested negative, while those with viral loads of
1E+03 copies/ml gave positive results in about half of samples.
CONCLUSIONS
The results obtained in this study are similar to datasets submitted by QCMD participants. All three
methods detect equally high viral loads in DBS but they should be improved to identify also lower
ones. The “CMV, HHV6, 7, 8 R-gene” kit seems to be particularly interesting, as it provides a readyto-
use and cheap mix which permits also the simultaneous detection of different herpesviruses.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Keywords:
CMV ; Cytomegalovirus ; DBS ; Dried Blood Spot ; CMV-DNA ; QCMD
Elenco autori:
L. Bubba, A. Mammoliti, S. Binda, M. Gambino, L. Pellegrinelli, V. Primache, M. Barbi
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