Quantitation of genomic DNA in plasma and serum samples: higher concentrations of genomic DNA found in serum than in plasma

Lee, Tzong‐Hae; Montalvo, Leilani; Chrebtow, Vera; Busch, Michael P.

doi:10.1046/j.1537-2995.2001.41020276.x

Cited by 273 publications

(165 citation statements)

References 21 publications

(39 reference statements)

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“…In addition, we were reassured by the results showing significantly more cell-free DNA in male sera compared with female sera, consistent with reports from other research groups (24). A study (27) pointed to finding more cell-free DNA in the serum in contrast to plasma due to lysis of white blood cells during centrifugation in the extraction step. The present study compared relative rather than absolute concentrations avoiding the white blood cell confounding factor.…”

Section: Discussionsupporting

confidence: 89%

Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology

Hart

Patton

Jacobson

et al. 2005

J Assist Reprod Genet

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Purpose : DNA-damaging factors have been reported in patients that failed to achieve pregnancy after assisted reproductive technologies (ART). The hypothesis was that increased circulating cell-free DNA released by damaged cells could predict unfavorable conditions leading to failed ART treatment. The objective was to compare the relative concentrations of cell-free DNA in the luteal phase sera of nonpregnant versus pregnant patients. Methods : Frozen-thawed sera (30 IVF cases) were obtained 1 week after embryo transfer. There were 16 pregnant and 14 nonpregnant cases and controls consisting of male sera (n = 8 cases). Modified isocratic capillary electrophoresis was performed and the images analyzed for cell-free DNA. Results : Circulating cell-free DNA were identified in the sera of all patients. The serum concentrations of high (12 kb) and low (1 kb) molecular weight cell-free DNA were similar for both nonpregnant and pregnant patients. Male control sera had higher cell-free DNA concentrations compared with females. Evaluation of sera from a control case showed no fluctuations in cell-free DNA concentrations throughout specific days of the menstrual cycle. Conclusions : The results do not support the use of the luteal phase cell-free DNA concentration as a marker for failed pregnancies. The equal concentrations of high and low molecular weight cell-free DNA and ladder band-like gel patterns suggested cell apoptosis as the source of DNA.

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Section: Discussionsupporting

confidence: 89%

Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology

Hart

Patton

Jacobson

et al. 2005

J Assist Reprod Genet

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“…Two latter studies and their results were quite similar to our work, although our study differed in that we used fresh and frozen serum /plasma from the maternal blood of Haemophilia A carriers and we also used 2 pairs of primers for the SRY gene to obtain a reliable signal for fetal gender in a Real-Time duplex PCR analysis and finally, we used SYBR Green instead of a probe in this analysis. We were able to obtain a high degree of accuracy with fresh serum, rather than with plasma, which is consistence with several studies indicated that fetal DNA in serum is present in greater quantities than in plasma and its concentration increases some 100 times when samples are kept at 4°C for 4 to 5 days [36,37]. In case of the presence of the large amounts of non-target DNA which is the major problem related to amplifying low copy number DNA target sequences [38], we were well able to overcome this problem by combining duplex/ nested-PCR and LC-qPCR.…”

Section: Methodssupporting

confidence: 79%

Evaluation of an Improved Non- invasive Fetal Sex Determination in Haemophilia A Patients

Mokarizadeh

Mesbah‐Namin

2015

JCDR

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Background: Haemophilia A (HA) is the most severe sex-linked bleeding disorder that is characterized with non-controlled and often threatening Haemorrhage. Routine fetal sex determination in early pregnancy with Haemophilia is based on invasive procedures that can be dangerous to the mother and fetus.Aim: The goal of this study is to present an improved assay for the non-invasive fetal sex determination using a Real-Time duplex PCR on the free fetal DNA (ffDNA) obtained from the maternal serum of the HA carriers.

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“…Serum samples have been shown to contain cell free DNA generated during the clotting process by the in vitro lysis of white blood cells. 34 Also for samples obtained from melanoma patients, the method of blood processing has been shown to significantly affect the levels of circulating DNA extracted. 35 Our data comparing DNA yields from plasma and from serum in matched samples from healthy subjects indicated that higher levels are recovered from serum samples under our experimental conditions.…”

Section: Discussionmentioning

confidence: 99%

Detection of mutated BRAFV600E variant in circulating DNA of stage III–IV melanoma patients

Daniotti

Vallacchi

Rivoltini

et al. 2007

Intl Journal of Cancer

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BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell‐free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow‐up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild‐type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I–II; half of the positives were obtained presurgery and half at follow‐up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma. © 2007 Wiley‐Liss, Inc.

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Quantitation of genomic DNA in plasma and serum samples: higher concentrations of genomic DNA found in serum than in plasma

Cited by 273 publications

References 21 publications

Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology

Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology

Evaluation of an Improved Non- invasive Fetal Sex Determination in Haemophilia A Patients

Detection of mutated BRAFV600E variant in circulating DNA of stage III–IV melanoma patients

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