Untitled

Serdyuk, O. I.; Ботезату, И. В.; Шелепов, В. П.; Потапова, Г. И.; Alekhina, R. P.; Molyaka, Yu. K.; Ананьев, В. С.; Кныш, В И; Melkonyan, O. S.; Umanskiĭ, S R; Lichtenshtein, A. V.

doi:10.1023/a:1017624120807

Cited by 10 publications

(1 citation statement)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Urine has the potential to serve as a non-invasive sampling method for the diagnosis of human disease and for molecular epidemiology. Numerous studies over the last decade have evaluated the diagnostic utility of urine with sometimes inconsistent results [18–21, 41–50]. This is in part due to the lack of standardized techniques for collecting, preserving, purifying and amplifying nucleic acids from urine.…”

Section: Discussionmentioning

confidence: 99%

An efficient and cost-effective method for purification of small sized DNAs and RNAs from human urine

Zainabadi¹,

Dhayabaran²,

Moideen³

et al. 2019

PLoS ONE

View full text Add to dashboard Cite

Urine holds great promise as a non-invasive sampling method for molecular diagnostics. The cell-free nucleic acids of urine however are small, labile, and difficult to purify. Here an efficient method for the purification of these nucleic acids is presented. An empirically derived protocol was devised by first identifying conditions that allowed recovery of a 100 base pair (bp) DNA, followed by optimization using a quantitative polymerase chain reaction (qPCR) assay. The resulting method efficiently purifies both small sized DNAs and RNAs from urine, which when combined with quantitative reverse transcription PCR (qRTPCR), demonstrably improves detection sensitivity. Fractionation experiments reveal that nucleic acids in urine exist both in the cell-free and cellular fraction, roughly in equal proportion. Consistent with previous studies, amplicons > 180bp show a marked loss in PCR sensitivity for cell-free nucleic acids. Finally, the lysis buffer developed here also doubles as an effective preservative, protecting against nucleic acid degradation for at least two weeks under simulated field conditions. With this method, volumes of up to 25ml of whole urine can be purified in a high-throughput and cost-effective manner. Coupled with its ability to purify both DNA and RNA, the described method may have broad applicability for improving the diagnostic utility of urine, particularly for the detection of low abundant targets.

show abstract

Section: Discussionmentioning

confidence: 99%

An efficient and cost-effective method for purification of small sized DNAs and RNAs from human urine

Zainabadi¹,

Dhayabaran²,

Moideen³

et al. 2019

PLoS ONE

View full text Add to dashboard Cite

show abstract

Molekulare Diagnostik in der urologischen Onkologie

et al. 2003

View full text Add to dashboard Cite

The goal of molecular diagnostics in oncology is the early diagnosis of malignant disease processes during initial work-up or as part of follow-up. Body fluids serve as the primary material for non-invasive diagnostic methods. Besides actual tumor cells, the examination of urine can yield evidence of secreted proteins or even free nucleic acids. In principle, all of the methods available for the detection of tumor markers in tissue or blood samples can be successfully applied to the examination of urine samples. However, molecular biological examination of urine samples is associated with important problems because the cells in such samples are exposed to significant degradation and regression effects and because certain components of the urine act to inhibit the polymerase chain reaction. The present overview discusses the respective strengths and weakness of the available technology as applied to the diagnosis of urologic malignancies. Experimental studies conducted to date have reported high sensitivities and specificities for molecular diagnostics using urine samples. It is important to note that not only carcinomas of the urinary bladder can be diagnosed from material obtained in urine samples: in fact, the method can be used to diagnose entities such as renal cell and prostate carcinomas and, due to renal filtration of DNA, even non-urologic malignancies. The diagnostic application of these methods, however, remains in an experimental stage and must still clear several hurdles before becoming available for routine clinical use.

show abstract