Implications of storing urinary DNA from different populations for molecular analyses

Angela Cannas, Glendah Kalunga, Clare Green, Ludovica Calvo, Patrick Katemangwe, Klaus Reither, Mark D. Perkins, Leonard Maboko, Michael Hoelscher, Elizabeth A. Talbot, Peter Mwaba, Alimuddin I. Zumla, Enrico Girardi, Jim F. Huggett

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Article numbere6985
JournalPLoS One
Volume4
Issue number9
DOIs
Publication statusPublished - Sep 10 2009

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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