Recovery of nanogram quantities of DNA from plasma and quantitative measurement using labeling by nick translation

Fournié, Gilbert J.; Gayral-Taminh, Martine; Bouché, Jean‐Pierre; Conté, J

doi:10.1016/0003-2697(86)90545-2

Cited by 26 publications

(11 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This work, conducted with a new method of DNA quantitation [3], confirms and extends the finding that plasma DNA concentration increases during hemodialy sis, reaching high levels at the end of the session [1]. Methodological differences from previous methods [8,9] may explain why plasma DNA levels have been found lower than initially described.…”

Section: Discussionsupporting

confidence: 66%

“…Until now, due to the lack of a suitable method of quantitation of plasma DNA, this hypothesis has not been challenged, and the physiopathological significance of this phenomenon has not been investigated. A new method of quantitation of plasma DNA based on the microextraction of DNA from plasma and on its internal labelling using a nick transla tion reaction has been set up [3]. In the present study, it has been applied to the serial measurement of plasma DNA levels in patients undergoing hemodialysis and hemofiltration and to the comparison of levels found at the same time in arterial and venous lines during the sessions.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Plasma DNA in Patients Undergoing Hemodialysis or Hemofiltration: Cytolysis in Artificial Kidney Is Responsible for the Release of DNA in Circulation

et al. 1989

View full text Add to dashboard Cite

Levels of circulating DNA increase under treatment by an artificial kidney. Using a new assay, levels of plasma DNA are studied in 45 patients during 99 sessions of hemodialysis or hemofiltration. Before the session, plasma DNA levels are increased in 41/99 samples and, among them, in 18/24 samples collected from hepatitis B surface antigen carriers. During the first 3 h of the session, plasma DNA levels increase whatever the method of treatment. At the 30th and 60th minute of hemodialysis, a positive gradient of plasma DNA exists between the output and the input of the artificial kidney. It is concluded that: (1) the increase in plasma DNA is related to the overall procedure of artificial kidney therapy; (2) death of leukocytes in the artificial kidney is responsible for the release and the increase in circulation of extracellular DNA.

show abstract

Section: Discussionsupporting

confidence: 66%

Section: Introductionmentioning

confidence: 99%

Plasma DNA in Patients Undergoing Hemodialysis or Hemofiltration: Cytolysis in Artificial Kidney Is Responsible for the Release of DNA in Circulation

et al. 1989

View full text Add to dashboard Cite

show abstract

“…4°C), half of the supernatant plasma was collected and stored at -20°C until used. DNA was measured in a few microliter of plasma using a radioisotopic method [10]. For reference, DNA assays were conducted twice at a weekly interval in 10 normal subjects aged 23-47 years (mean ± SD: 29 ± 9); similar results were obtained on DO and D7 (mean ± SD: 9 ± 5 and 8 ± 4 ng/ml, respectively).…”

Section: Dna Assaymentioning

confidence: 63%

Plasma DNA as Cell Death Marker in Elderly Patients

et al. 1993

Self Cite

View full text Add to dashboard Cite

Plasma DNA increases where cell death occurs in vivo. To investigate its significance in elderly patients, plasma DNA was assayed in 79 institutionalized patients over 68 years of age. The patients were divided into two groups: group I comprises 39 patients suffering from various acute or chronic illnesses; group II comprises 40 patients without chronic disease, and free of any clinical or biological symptoms of any infectious or inflammatory process. Plasma DNA was higher in group I than in group II (p < 0.0001) and in group II than in a control group of middle-aged subjects (p < 0.05). In group I, increase in plasma DNA concentration was found in various pathological situations associated with cell death phenomena, including infections, cancers with metastasis, hepatitis, irreversible cardiac failure, severe respiratory insufficiency and thrombophlebitis. Plasma DNA concentrations were not correlated with erythrocyte sedimentation rate, fibrinogen concentration, hemoglobin concentration or leukocyte count. In group I, as well as in the overall population, survival after 1 month was significantly reduced in patients with increased concentrations of plasma DNA. In conclusion, plasma DNA as a marker of cell death phenomena occurring in vivo, could be helpful for follow-up and management of elderly patients.

show abstract

“…For plasma preparation, EDTA-coated or citrate Vacutainer tubes were used to avoid the potential inhibitory effects of heparin on the amplification assay (Beutler et al, 1990 In extraction method two (six patients, 15 normal volunteers), plasma or serum DNA was co-precipitated with gelatin by a method modified from that of Fournie et al (1986). A stock 5% (w/v) gelatin solution was prepared by mixing 1 g of gelatin (G8-500, Fisher, Pittsburgh, PA, USA) with 20 pl of sterile, doubledistilled water, autoclaving for 30 min and filtering through a 0.2-gm filter.…”

Section: Specimensmentioning

confidence: 99%

Detection of mutant K-ras DNA in plasma or serum of patients with colorectal cancer

Kopreski

Benko²,

Kwee³

et al. 1997

Br J Cancer

145

View full text Add to dashboard Cite

Summary Increased understanding of the molecular basis of colorectal cancer and recognition that extracellular DNA circulates in the plasma and serum of cancer patients enables new approaches to detection and monitoring. We used a polymerase chain reaction (PCR) assay to demonstrate mutant K-ras DNA in the plasma or serum of patients with colorectal cancer. Plasma or serum was fractionated from the blood of 31 patients with metastatic or unresected colorectal cancer and from 28 normal volunteers. DNA was extracted using either a sodium chloride or a gelatin precipitation method and then amplified in a two-stage PCR assay using selective restriction enzyme digestion to enrich for mutant K-ras DNA. Mutant K-ras DNA was detected in the plasma or serum of 12 (39%) patients, all confirmed by sequencing, but was not detected in any of the normal volunteers. K-ras mutations were detected in plasma or serum regardless of sex, primary tumour location, principal site of metastasis or proximity of chemotherapy and surgery to blood sampling. Tumour specimens available for 19 of the patients were additionally assayed for ras mutations and compared with blood specimens. Our results indicate mutant K-ras DNA is readily detectable by PCR in the plasma or serum of patients with advanced colorectal cancer. Thus, plasma-or serum-based nucleic acid amplification assays may provide a valuable method of monitoring and potentially detecting colorectal cancer.

show abstract

Recovery of nanogram quantities of DNA from plasma and quantitative measurement using labeling by nick translation

Cited by 26 publications

References 17 publications

Plasma DNA in Patients Undergoing Hemodialysis or Hemofiltration: Cytolysis in Artificial Kidney Is Responsible for the Release of DNA in Circulation

Plasma DNA in Patients Undergoing Hemodialysis or Hemofiltration: Cytolysis in Artificial Kidney Is Responsible for the Release of DNA in Circulation

Plasma DNA as Cell Death Marker in Elderly Patients

Detection of mutant K-ras DNA in plasma or serum of patients with colorectal cancer

Contact Info

Product

Resources

About