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Rapid detection and quantification of CMV DNA in urine using LightCycler (TM)-based real-time PCR

Lookup NU author(s): Dr Andrew Turner


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A real-time quantitative PCR-hybridisation assay was developed for the detection of human cytomegalovirus DNA in clinical material. The assay is based on a LightCycler (LC) and provides both rapid results (< 1 h) and quantification over a broad dynamic range (2 x 10(3)-5 x 10(8) CMV DNA copies/ml). Given that the assay showed a 3-fold increase in sensitivity compared to detection of early antigen fluorescent foci (DEAFF) testing of urine samples, we investigated the practicality of testing surveillance such specimens from immunocompromised patients at risk of CMV disease. Over a 12-month period, CMV DNA was detected in 81 (7%) of 1154 urine samples examined. A total of 28 patients tested positive; urine viral loads were higher in 13 infants being investigated for suspected congenital infection (median 1.6 x 10(5) copies/ml) compared with 15 transplant recipients (median 9 x 10(3) copies/ml). Urine samples could be tested directly without processing such that results were available in < 1h. Real-time PCR provided information on the quantification of CMV DNA in urine and proved a reliable method for the surveillance of immunocompromised patients at risk of CMV disease. This approach should facilitate a better understanding of the epidemiology and natural history of CMV disease. Moreover, LC-based quantitative PCR is a potentially valuable tool for the management of CMV disease; assisting with the prompt initiation of treatment and assessing therapeutic response. (C) 2002 Elsevier Science B.V. All rights reserved.

Publication metadata

Author(s): Kearns AM, Turner AJL, Eltringham GJA, Freeman R

Publication type: Article

Publication status: Published

Journal: Journal of Clinical Virology

Year: 2002

Volume: 24

Issue: 1-2

Pages: 131-134

ISSN (print): 1386-6532

ISSN (electronic): 1873-5967

Publisher: Elsevier BV


DOI: 10.1016/S1386-6532(01)00240-2


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