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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Implications of storing urinary DNA from different populations for molecular analyses
PLoS ONE, Volume 4, No. 9, Article e6985, Year 2009
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Description
Background: Molecular diagnosis using urine is established for many sexually transmitted diseases and is increasingly used to diagnose tumours and other infectious diseases. Storage of urine prior to analysis, whether due to home collection or bio-banking, is increasingly advocated yet no best practice has emerged. Here, we examined the stability of DNA in stored urine in two populations over 28 days. Methodology: Urine from 40 (20 male) healthy volunteers from two populations, Italy and Zambia, was stored at four different temperatures (RT, 4°C, -20°/C & -80°C) with and without EDTA preservative solution. Urines were extracted at days 0, 1, 3, 7 and 28 after storage. Human DNA content was measured using multi-copy (ALU J) and single copy (TLR2) targets by quantitative real-time PCR. Zambian and Italian samples contained comparable DNA quantity at time zero. Generally, two trends were observed during storage; no degradation, or rapid degradation from days 0 to 7 followed by little further degradation to 28 days. The biphasic degradation was always observed in Zambia regardless of storage conditions, but only twice in Italy. Conclusion: Site-specific differences in urine composition significantly affect the stability of DNA during storage. Assessing the quality of stored urine for molecular analysis, by using the type of strategy described here, is paramount before these samples are used for molecular prognostic monitoring, genetic analyses and disease diagnosis. © 2009 Cannas et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC2735781/bin/pone.0006985.s001.xls
https://efashare.b-cdn.net/share/pmc/articles/PMC2735781/bin/pone.0006985.s002.doc
https://efashare.b-cdn.net/share/pmc/articles/PMC2735781/bin/pone.0006985.s003.doc
https://efashare.b-cdn.net/share/pmc/articles/PMC2735781/bin/pone.0006985.s004.doc
Authors & Co-Authors
Cannas, Angela
Italy, Rome
Irccs Istituto Nazionale Malattie Infettive Lazzaro Spallanzani
Kalunga, Glendah
Zambia, Lusaka
University Teaching Hospital Lusaka
Green, Clare
Zambia, Lusaka
University Teaching Hospital Lusaka
United Kingdom, London
University College London
Calvo, Ludovica
Italy, Rome
Irccs Istituto Nazionale Malattie Infettive Lazzaro Spallanzani
Katemangwe, Patrick
Zambia, Lusaka
University Teaching Hospital Lusaka
Reither, Klaus
Tanzania
Nimr-mbeya Medical Research Programme
Germany, Munich
Klinikum Der Universität München
Perkins, Mark D.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Maboko, Leonard L.
Tanzania
Nimr-mbeya Medical Research Programme
Höelscher, Michael
Tanzania
Nimr-mbeya Medical Research Programme
Germany, Munich
Klinikum Der Universität München
Talbot, Elizabeth A.S.
Switzerland, Geneva
Foundation for Innovative new Diagnostics, Switzerland
Mwaba, Peter B.
Zambia, Lusaka
University Teaching Hospital Lusaka
Zumla, A. I.
Zambia, Lusaka
University Teaching Hospital Lusaka
United Kingdom, London
University College London
Girardi, Enrico
Italy, Rome
Irccs Istituto Nazionale Malattie Infettive Lazzaro Spallanzani
Huggett, Jim
Zambia, Lusaka
University Teaching Hospital Lusaka
United Kingdom, London
University College London
Statistics
Citations: 51
Authors: 14
Affiliations: 6
Identifiers
Doi:
10.1371/journal.pone.0006985
e-ISSN:
19326203
Research Areas
Genetics And Genomics
Study Approach
Quantitative
Study Locations
Zambia
Participants Gender
Male