1. Evaluation of the Automated QIAsymphony SP/AS Workflow for Cytomegalovirus DNA Extraction and Amplification from Dried Blood Spots
- Author
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Daniela Spalletti-Cernia, Caterina Rocco, Luca Vallefuoco, Roberta De Mattia, Rosanna Sorrentino, Pasquale Di Costanzo, Sara Barbato, Claudia Mazzarella, Francesco Raimondi, Antonella Giannattasio, Giuseppe Portella, Spalletti, CERNIA DANIELA, Barbato, Sara, Sorrentino, Rosanna, Vallefuoco, Luca, Rocco, Caterina, Di Costanzo, Pasquale, Giannattasio, Antonella, Raimondi, Francesco, Mazzarella, Claudia, DE MATTIA, Roberta, and Portella, Giuseppe
- Subjects
0301 basic medicine ,Human cytomegalovirus ,Cytomegalovirus Infection ,Male ,030106 microbiology ,Congenital cytomegalovirus infection ,Cytomegalovirus ,Biology ,Real-Time Polymerase Chain Reaction ,Sensitivity and Specificity ,law.invention ,Workflow ,03 medical and health sciences ,Automation ,0302 clinical medicine ,law ,Virology ,medicine ,Humans ,030212 general & internal medicine ,Polymerase chain reaction ,Newborn screening ,Infant, Newborn ,virus diseases ,Cytomegaloviru ,Viral Load ,medicine.disease ,DNA extraction ,Dried blood spot ,Infectious Diseases ,Real-time polymerase chain reaction ,Immunology ,Cytomegalovirus Infections ,DNA, Viral ,Female ,Dried Blood Spot Testing ,Viral load ,Human - Abstract
Objective: Human cytomegalovirus (CMV) can be considered the most important agent of congenital infection. Long-term sequelae of congenital infection occur in about 15% of infants asymptomatic at birth. To avoid long-term sequelae or to reduce their burden, it is necessary to identify infected children for early interventions. CMV DNA can be detected in dried blood spots (DBSs). DBSs have been used in several studies for the retrospective diagnosis of congenital CMV (CCMV). It has been proposed to use DBSs for the newborn screening of CMV infection; however, manual methods are not suitable for newborn screening of CCMV. Methods: We evaluated in an off-label application the use of an automated instrument, the QIAsymphony SP/AS, in combination with the artus CMV QS-RGQ kit and the RotorGene Q real-time polymerase chain reaction system. Results: We analyzed 100 DBSs from newborns positive or negative for plasma CMV DNA with a 94% concordance in positive samples. Conclusions: We show that the QIAsymphony SP/AS and RotorGene Q workflow is suitable for CMV DNA extraction and detection from DBSs and that the system correctly identified newborns at risk of late sequelae due to CMV infection.
- Published
- 2016