Influence of delay in blood-processing on the level of cell-free fetal DNA in maternal plasma sample
Journal Title: Di-er Junyi Daxue Xuebao - Year 2008, Vol 29, Issue 9
Abstract
Objective:To investigate the influencing factors of cell-free fetal DNA level in the maternal plasma during blood-processing.Methods: Aliquots of blood samples from pregnant women with male fetus were processed at 6 h and 36 h after sampling.The SRY and β-globin genes and the total DNA level were quantified by real-time quantitative PCR.Death of white blood cells was assayed by flow cytometry after stained with AnnexinⅤ/PI.The plasma DNase activity was assayed by radial enzyme-diffusion method and plasma lactate dehydrogenases(LDH) by rate method.Results: A 36 hour delay in blood-processing led to a significant increase in the total DNA and decrease in the fetal DNA (SRY gene) in the maternal plasma.The ratio of fetal DNA decreased from (10.3±5.6)% at 6 h after sampling to (3.0±2.1)% at 36 h after sampling under 4℃(P<0.05).No dead cells were identified in the blood sample 6 h after sampling; however,apoptosis and necrosis of white blood cells were identified 36 h after sampling.The activity of LDH at 36 h was significantly higher than that at 6 h (P<0.05).Radial enzyme-diffusion result showed that,though greatly decreased at 4℃,the DNase was still able to degrade DNA.Conclusion: Delay in blood-processing can lead to increase of the total free DNA in maternal plasma but decrease of fetal DNA,which might be related to the death of white blood cells and degradation of fetal DNA by plasma DNase,so the extraction of fetal DNA should be done as early as possible after sampling (within 6 h).
Authors and Affiliations
Qin LI
Keywords
Related Articles
- EP ID EP84208
- DOI -
- Views 152
- Downloads 0
Pharmacokinetics of granisetron hydrocloride capsule in healthy volunteers
Objective:To establish a sensitive and specific LC/MS/MS method for the determination of granisetron in human plasma,and to apply it in investigating the pharmacokinetics of granisetron hydrocloride capsule in healthy vo...
LC-MS/MS in determination of telmisartan concentration in human plasma
Influence of Traditional Chinese Recipe Xiaotanjieyu Decoction on behavior of mice after chronic swimming stress and on expression of 5HT1A receptor mRNA in their hippocampus
5α-双氢睾酮对前列腺癌LNCaP细胞钙离子移动和细胞生长的影响
目的:探讨5α-双氢睾酮(DHT)对前列腺癌LNCaP细胞钙离子移动的影响及其机制。方法:应用Fura-2/AM Ca2+ 荧光探针法结合MiraCal荧光成像系统动态检测DHT刺激以及Ca2+通道阻滞剂干预后LNCaP细胞内钙离子浓度([Ca2+]i)的变化。应用MTT法观察细...
HPLC法同时测定中药丹参中水溶性和脂溶性成分的含量
目的:建立同时测定丹参中2种水溶性成分和2种脂溶性成分的方法。方法:HPLC-DAD法,采用Agilent Zorbax TC C18柱(4.6 mm×250 mm,5 μm),以甲醇2%醋酸为流动相进行梯度洗脱:0～15 min,30% B～40% B; 15～20 min,40% B～60% B; 20～25 min,60%...