kjlm The Korean Journal of Laboratory Medicine Korean J Leg Med 2383-5702 2383-5710 The Korean Society for Legal Medicine 10.7580/kjlm.2016.40.3.78 kjlm-40-78 Original Article Factors Affecting DNA Yields from Serum and Plasma Samples Used for Personal Identification Testing LeeJi Hyun 1∗ KimBoram 1∗ ChoSohee 2 SeoHee Jin 1 LeeSoong Deok 1 2 Department of Forensic Medicine, Seoul National University College of Medicine, Seoul, Korea Institute of Forensic Science, Seoul National University College of Medicine, Seoul, Korea Correspondence to Soong Deok Lee Department of Forensic Medicine, Seoul National University College of Medicine, 103 Daehak-ro, Jongno-gu, Seoul 03080, Korea Tel: +82-2-740-8359 Fax: +82-2-764-8340 E-mail: sdlee@snu.ac.kr

These authors contributed equally to this work.

 122016 19122016 403 7882 26072016 22082016 26082016 Copyright © 2016 The Korean Society for Legal Medicine 2016 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Serum or plasma is free of cellular components. As DNA is in the nucleus or mitochondria of a cell, it can be presumed that serum/plasma is DNA free. However, there are cases wherein serum/plasma is the only resource available for identification analysis, yet no sufficient data are available regarding whether reliable DNA testing can be applied to such cases, and what the influencing factors are when testing is a valid course of action. The aim of this study is to illustrate the factors that can be used in the genetic testing of serum/plasma when identifying an individual. The results showed that the concentration of serum DNA significantly increased over time in 4°C storage, and the DNA yields from samples stored in heparin tubes were overall higher than from samples stored in ethylenediaminetetraacetic acid tubes. We observed that the concentration of DNA in serum successfully matched 100% to the short tandem repeat data of blood DNA.

 Serum Plasma DNA fingerprinting Identification REFERENCES Lo YM Tein MS Lau TK Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis Am J Hum Genet 199862 76875 Lee JH Lee HY Cho S DNA profiling via short tandem repeat analysis by using serum samples Korean J Leg Med 201337 2203 Takayama T Yamada S Watanabe Y Origin of DNA in human serum and usefulness of serum as a material for DNA typing Leg Med (Tokyo) 20013 10913 Honda H Miharu N Ohashi Y Fetal gender determination in early pregnancy through qualitative and quantitative analysis of fetal DNA in maternal serum Hum Genet 2002110 759 Bischoff FZ Sinacori MK Dang DD Cell-free fetal DNA and intact fetal cells in maternal blood circulation: implications for first and second trimester non-invasive prenatal diagnosis Hum Reprod Update 20028 493500 Allen RW Pritchard JK Resolution of a serum sample mix-up through the use of short tandem repeat DNA typing Transfusion 200444 17504 Lam NY Rainer TH Chiu RW EDTA is a better anticoagulant than heparin or citrate for delayed blood processing for plasma DNA analysis Clin Chem 200450 2567 Ravard-Goulvestre C Crainic K Guillon F Successful extraction of human genomic DNA from serum and its application to forensic identification J Forensic Sci 200449 603 Barra GB Santa Rita TH Chianca CF Fetal male lineage determination by analysis of Y-chromosome STR haplotype in maternal plasma Forensic Sci Int Genet 201515 10510 Sedlackova T Repiska G Celec P Fragmentation of DNA affects the accuracy of the DNA quantitation by the commonly used methods Biol Proced Online 201315 5 Figures and Tables

Box plots of DNA concentrations in serum (A, B) and plasma (C). RT, room temperature; EDTA, ethylenediaminetetraacetic acid.

Electropherograms of the amplified DNA from one sample. (A) Peak exceeding the 50-RFU threshold is present. (B) Peaks exceeding the 40-RFU threshold are present. RFU, relative fluorescence unit.

Amount of DNA recovered from donors (15 and 14) with time-temperature differences by two-way ANOVA

Time RT (ng/μL) 4°C (ng/μL) t p-value
0 hr 0.1441±0.1480 0.1441±0.1480 0 >0.990
1 hr 0.1525±0.1228 0.1875±0.2687 0.0799 0.937
2 hr 0.1125±0.1136 0.0839±0.0723 0.0652 0.948
4 hr 0.1039±0.0778 0.2597±0.2171 0.3554 0.723
8 hr 0.1636±0.3502 0.2216±0.1060 0.1323 0.895
12 hr 0.1530±0.2022 0.3362±0.2755 0.4179 0.677
24 hr 0.1177±0.0682 0.9903±0.9097 1.991 0.049
48 hr 0.2321±0.1585 4.939±4.767 10.74 <0.001
0 day 0.2025±0.1938 0.2025±0.1938 0 >0.990
1 day 0.0552±0.0338 0.7881±1.099 0.8800 0.380
2 days 0.0639±0.0561 1.845±2.136 2.138 0.033
3 days 0.1391±0.1136 3.228±3.150 3.709 <0.001
4 days 0.1746±0.1018 2.961±3.026 3.345 0.001
5 days 0.3432±0.1755 3.030±3.250 3.226 0.001
6 days 0.5806±0.2739 3.674±5.452 3.714 <0.001
7 days 0.5105±0.2885 2.798±2.936 2.746 0.006
8 days 0.7600±0.3676 2.768±4.107 2.411 0.017
9 days 1.066±0.5695 2.144±3.483 1.295 0.196
10 days 1.444±0.6514 1.670±1.706 0.2717 0.786

Values are presented as mean±standard deviation. RT, room temperature.

Amount of DNA recovered from 15 donors with time-anticoagulant (heparin, EDTA) differences by two-way ANOVA

Time Heparin (ng/μL) EDTA (ng/μL) t p-value
0 hr 0.2672±0.1352 0.0448±0.0661 2.608 0.013
1 hr 0.8045±0.3414 0.0877±0.0828 8.406 <0.001
3 hr 0.0712±0.1836 0.2056±0.0621 1.576 0.123
7 hr 0.4122±0.1797 0.1632±0.4480 2.920 0.006

Values are presented as mean±standard deviation. EDTA, ethylenediaminetetraacetic acid.