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Filtration Device for On-Site Collection, Storage and Shipment of Cells from Urine and Its Application to DNA-Based Detection of Bladder Cancer.

Andersson E, Dahmcke CM, Steven K, Larsen LK, Guldberg P - PLoS ONE (2015)

Bottom Line: In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%).Only one patient in this group was positive for a FGFR3 mutation.This patient had a stage Ta tumor resected 6 months later.

View Article: PubMed Central - PubMed

Affiliation: Danish Cancer Society Research Center, Copenhagen, Denmark.

ABSTRACT
Molecular analysis of cells from urine provides a convenient approach to non-invasive detection of bladder cancer. The practical use of urinary cell-based tests is often hampered by difficulties in handling and analyzing large sample volumes, the need for rapid sample processing to avoid degradation of cellular content, and low sensitivity due to a high background of normal cells. We present a filtration device, designed for home or point-of-care use, which enables collection, storage and shipment of urinary cells. A special feature of this device is a removable cartridge housing a membrane filter, which after filtration of urine can be transferred to a storage unit containing an appropriate preserving solution. In spiking experiments, the use of this device provided efficient recovery of bladder cancer cells with elimination of >99% of excess smaller-sized cells. The performance of the device was further evaluated by DNA-based analysis of urinary cells collected from 57 patients subjected to transurethral resection following flexible cystoscopy indicating the presence of a tumor. All samples were tested for FGFR3 mutations and seven DNA methylation markers (BCL2, CCNA1, EOMES, HOXA9, POU4F2, SALL3 and VIM). In the group of patients where a transitional cell tumor was confirmed at histopathological evaluation, urine DNA was positive for one or more markers in 29 out of 31 cases (94%), including 19 with FGFR3 mutation (61%). In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%). Only one patient in this group was positive for a FGFR3 mutation. This patient had a stage Ta tumor resected 6 months later. The ability to easily collect, store and ship diagnostic cells from urine using the presented device may facilitate non-invasive testing for bladder cancer.

No MeSH data available.


Related in: MedlinePlus

Enrichment of bladder tumor cells from urine.Urine samples from patients with bladder tumors were divided into two fractions and processed by device filtration and sedimentation, respectively. Equimolar amounts of DNA from filters and sediments were tested for FGFR3 mutations using ddPCR.
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pone.0131889.g004: Enrichment of bladder tumor cells from urine.Urine samples from patients with bladder tumors were divided into two fractions and processed by device filtration and sedimentation, respectively. Equimolar amounts of DNA from filters and sediments were tested for FGFR3 mutations using ddPCR.

Mentions: In order to test whether filtration could increase the ratio of normal-to-tumor cells, we first tested 13 urine samples in a split-sample setup, where 25 ml of each sample were sedimented by centrifugation, and the remainder was processed by filtration. DNA isolated from all filter and sediment samples were screened for four common FGFR3 mutations (p.R248C, p.S249C, p.G370C and p.Y373C) using ddPCR. Eight of the samples (58%) were positive for one of these mutations (Table 1). Quantitative analysis using equimolar amounts of total DNA from filters and sediments showed that the ratio of mutant-to-wild type DNA was higher in the filtered samples than in the corresponding sediments (Table 3). Most important, the greatest enrichments (6.5 and 8.0 times, respectively) were achieved for the two samples representing the lowest mutant-to-wild type ratios (Fig 4).


Filtration Device for On-Site Collection, Storage and Shipment of Cells from Urine and Its Application to DNA-Based Detection of Bladder Cancer.

Andersson E, Dahmcke CM, Steven K, Larsen LK, Guldberg P - PLoS ONE (2015)

Enrichment of bladder tumor cells from urine.Urine samples from patients with bladder tumors were divided into two fractions and processed by device filtration and sedimentation, respectively. Equimolar amounts of DNA from filters and sediments were tested for FGFR3 mutations using ddPCR.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495058&req=5

pone.0131889.g004: Enrichment of bladder tumor cells from urine.Urine samples from patients with bladder tumors were divided into two fractions and processed by device filtration and sedimentation, respectively. Equimolar amounts of DNA from filters and sediments were tested for FGFR3 mutations using ddPCR.
Mentions: In order to test whether filtration could increase the ratio of normal-to-tumor cells, we first tested 13 urine samples in a split-sample setup, where 25 ml of each sample were sedimented by centrifugation, and the remainder was processed by filtration. DNA isolated from all filter and sediment samples were screened for four common FGFR3 mutations (p.R248C, p.S249C, p.G370C and p.Y373C) using ddPCR. Eight of the samples (58%) were positive for one of these mutations (Table 1). Quantitative analysis using equimolar amounts of total DNA from filters and sediments showed that the ratio of mutant-to-wild type DNA was higher in the filtered samples than in the corresponding sediments (Table 3). Most important, the greatest enrichments (6.5 and 8.0 times, respectively) were achieved for the two samples representing the lowest mutant-to-wild type ratios (Fig 4).

Bottom Line: In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%).Only one patient in this group was positive for a FGFR3 mutation.This patient had a stage Ta tumor resected 6 months later.

View Article: PubMed Central - PubMed

Affiliation: Danish Cancer Society Research Center, Copenhagen, Denmark.

ABSTRACT
Molecular analysis of cells from urine provides a convenient approach to non-invasive detection of bladder cancer. The practical use of urinary cell-based tests is often hampered by difficulties in handling and analyzing large sample volumes, the need for rapid sample processing to avoid degradation of cellular content, and low sensitivity due to a high background of normal cells. We present a filtration device, designed for home or point-of-care use, which enables collection, storage and shipment of urinary cells. A special feature of this device is a removable cartridge housing a membrane filter, which after filtration of urine can be transferred to a storage unit containing an appropriate preserving solution. In spiking experiments, the use of this device provided efficient recovery of bladder cancer cells with elimination of >99% of excess smaller-sized cells. The performance of the device was further evaluated by DNA-based analysis of urinary cells collected from 57 patients subjected to transurethral resection following flexible cystoscopy indicating the presence of a tumor. All samples were tested for FGFR3 mutations and seven DNA methylation markers (BCL2, CCNA1, EOMES, HOXA9, POU4F2, SALL3 and VIM). In the group of patients where a transitional cell tumor was confirmed at histopathological evaluation, urine DNA was positive for one or more markers in 29 out of 31 cases (94%), including 19 with FGFR3 mutation (61%). In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%). Only one patient in this group was positive for a FGFR3 mutation. This patient had a stage Ta tumor resected 6 months later. The ability to easily collect, store and ship diagnostic cells from urine using the presented device may facilitate non-invasive testing for bladder cancer.

No MeSH data available.


Related in: MedlinePlus