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Development of a novel single tube nested PCR for enhanced detection of cytomegalovirus DNA from dried blood spots

Atkinson, C; Emery, VC; Griffiths, PD; (2014) Development of a novel single tube nested PCR for enhanced detection of cytomegalovirus DNA from dried blood spots. Journal of Virological Methods , 196 pp. 40-44. 10.1016/j.jviromet.2013.10.029. Green open access

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Abstract

Newborn screening for congenital cytomegalovirus (CCMV) using dried blood spots (DBS) has been proposed because many developed countries have DBS screening programmes in place for other diseases. The aim of this study was to develop a rapid, single tube nested polymerase chain reaction (PCR) method for enhanced detection of CMV from DBS compared to existing (single target) real time PCRs. The new method was compared with existing real time PCRs for sensitivity and specificity. Overall sensitivity of the single target PCR assays in both asymptomatic and symptomatic infants with laboratory confirmed congenital CMV was 69% (CMV PCR or culture positive before day 21 of life). In contrast, the single tube nested assay had an increased sensitivity of 81% with100% specificity. Overall the assay detected CMV from a DBS equivalent to an original blood sample which contained 500 IU/ml. In conclusion this single tube nested methodology allows simultaneous amplification and detection of CMV DNA in 1.5 h removing the associated contamination risk of a two step nested PCR. Owing to its increased sensitivity, it has the potential to be used as a screening assay and ultimately allow early identification and intervention for children with congenital CMV.

Type: Article
Title: Development of a novel single tube nested PCR for enhanced detection of cytomegalovirus DNA from dried blood spots
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.jviromet.2013.10.029
Publisher version: https://doi.org/10.1016/j.jviromet.2013.10.029
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Congenital cytomegalovirus, Dried blood spot, Nested PCR, Newborn screening
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity
URI: https://discovery.ucl.ac.uk/id/eprint/10050567
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