지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.1
- 수록면
- 18 - 27 (10page)
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초록· 키워드
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Background: Circulating levels of cell-free DNA increase in many pathologic conditions. However, notable discrepancies in the quantitative analysis of cell-free DNA from a large number of laboratories have become a considerable pitfall, hampering its clinical application.
Methods: We designed a novel recombinant DNA fragment that could be applied as an internal standard in a newly developed and validated duplex real-time PCR assay for the quantitative analysis of total cell-free plasma DNA, which was tested in 5,442 healthy adults and 200 trauma patients.
Results: Compared with two traditional methods, this novel assay showed a lower detection limit of 0.1 ng/mL, lower intra- and inter-assay CVs, and higher accuracy in the recovery test. The median plasma DNA concentration of healthy males (20.3 ng/mL, n=3,092) was significantly higher than that of healthy females (16.1 ng/mL, n=2,350) (Mann-Whitney two-sample rank sum test, P<0.0001). The reference intervals of plasma DNA concentration were 0-45.8 ng/mL and 0-52.5 ng/mL for healthy females and males, respectively. The plasma DNA concentrations of the majority of trauma patients (96%) were higher than the upper normal cutoff values and were closely related to the corresponding injury severity scores (R2=0.916, P<0.0001).
Conclusions: This duplex real-time PCR assay with a new internal standard could eliminate variation and allow for more sensitive, repeatable, accurate, and stable quantitative measurements of plasma DNA, showing promising application in clinical diagnosis.
목차
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참고문헌 (37)
참고문헌 신청
Devonshire AS / 2014 / Towards standardisation of cell-free DNA measurement in plasma: controls for extraction efficiency, fragment size bias and quantification / Anal Bioanal Chem 406:6499~6512
Ramakers C / 2003 / Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data / Neurosci Lett 339:62~66
Xue X / 2009 / Optimizing the yield and utility of circulating cell-free DNA from plasma and serum / Clin Chim Acta 404:100~104
Zhang P / 2004 / A new quantitative method for circulating DNA level in human serum by capillary zone electrophoresis with laser-induced fluorescence detection / Electrophoresis 25:1823~1828
Fujii S / 2013 / A coupling system of capillary gel electrophoresis with inductively coupled plasma-mass spectrometry for the determination of double stranded DNA fragments / Metallomics 5:424~428
Ramakers C / 2003 / Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data / Neurosci Lett 339:62~66
Xue X / 2009 / Optimizing the yield and utility of circulating cell-free DNA from plasma and serum / Clin Chim Acta 404:100~104
Zhang P / 2004 / A new quantitative method for circulating DNA level in human serum by capillary zone electrophoresis with laser-induced fluorescence detection / Electrophoresis 25:1823~1828
Fujii S / 2013 / A coupling system of capillary gel electrophoresis with inductively coupled plasma-mass spectrometry for the determination of double stranded DNA fragments / Metallomics 5:424~428
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