[1]王洁敏,向 芃,顾乐怡.不同预处理方法对提取尿液脱落细胞mRNA质量的影响[J].现代检验医学杂志,2016,31(04):125-127,130.[doi:10.3969/j.issn.1671-7414.2016.04.035]
 WANG Jie-min,XIANG Peng,GU Le-yi.Effects of Different Pretreatment Methods on mRNA Quality and Quantity of Exfoliated Urinary Cells[J].Journal of Modern Laboratory Medicine,2016,31(04):125-127,130.[doi:10.3969/j.issn.1671-7414.2016.04.035]
点击复制

不同预处理方法对提取尿液脱落细胞mRNA质量的影响()
分享到:

《现代检验医学杂志》[ISSN:/CN:]

卷:
第31卷
期数:
2016年04期
页码:
125-127,130
栏目:
研究简报·实验技术
出版日期:
2016-08-10

文章信息/Info

Title:
Effects of Different Pretreatment Methods on mRNA Quality and Quantity of Exfoliated Urinary Cells
文章编号:
1671-7414(2016)04-125-04
作者:
王洁敏向 芃顾乐怡
上海交通大学医学院附属仁济医院肾脏科,上海 200127
Author(s):
WANG Jie-minXIANG PengGU Le-yi
Renal Division, Renji Hospital,Shanghai Jiaotong University School of Medicine,Shanghai200127,China
关键词:
慢性肾脏病 尿液脱落细胞 mRNA 足细胞
分类号:
R446.12
DOI:
10.3969/j.issn.1671-7414.2016.04.035
文献标志码:
A
摘要:
目的 探索合适的处理尿标本的方法获得尿脱落细胞mRNA。方法 比较尿液容量、尿液标本冻存时间、处理方式对提取尿液脱落细胞RNA的产量与质量的影响,并进行qPCR的试验。结果 40 ml尿液标本组及10 ml尿液标本组RNA总量均值分别为886.94±222.93 ng及211.22±62.01 ng,两组A260nm/A280nm比值分别为1.80±0.10和1.77±0.09; 尿液标本量与RNA总量相关(t=3.604,P=0.002),不同尿液标本量RNA质量差异无统计学意义(t=0.708,P值>0.05)。实时离心提取及冰冻保存尿液7天后提取两种预处理方法RNA总量分别为886.94±945.84 ng及881.50±829.02 ng,A260nm/A280nm比值分别为1.80±0.10及1.80±0.87; RNA总量与A260nm/A280nm比值差异无统计学意义(t=-0.221~0.085,P值均>0.05)。直接冰冻尿液标本与TRIzol保存沉渣细胞两种预处理方法RNA总量分别为637.81±525.24 ng及639.86±535.42 ng,A260nm/A280nm比值分别为1.84±0.13及1.83±0.96; RNA总量与A260nm/A280nm比值差异无统计学意义(t=-0.47~0.293,P值均>0.05)。尿脱落细胞mRNA中可以检测到足细胞的标记蛋白WT-1,podocin和synaptopodin的基因表达。结论 尿液标本量是提取脱落细胞RNA量的关键因素,-80℃低温冻存尿标本7天或Trizol预处理冻存均对提取尿液沉渣细胞RNA数量和质量无影响。
Abstract:
Objective To explore a proper method to obtainexfoliated cells mRNA for detection in urine specimen.Methods Urine volume,different pretreating ways and frozen storage time on urinary sample were used to collect mRNA.A260nm/A280nm and A260nm/A280nm ratio,as well as qPCR were performed.Results Amount of mRNA was determinded by urine volume,but neitherA260nm/A280nm or A260nm/A280nm ratio wasrealated with urine volume.Compared with real-time extraction process,freezing urine samples for 7 days showed no statistical difference(P>0.05)in total RNA and A260nm/A280nm or A260nm/A280nm ratio.There was no significant difference in total RNA and A260nm/A280nm or A260nm/A280nm ratio between direct freezing urine samples and Trizol-treated urine exfoliated cells.Podocyte marker including WT-1,podocin and synaptopodin mRNAs were detected in urine sediment cells.Conclusion Quantity of mRNA was determinded by urine volume.Directly freezing urinesamples at -80 ℃ for 7 days or Trizol-treating urine sediment cells have no effect on lowing quality and quantity of urinary sediment mRNA.

参考文献/References:

[1] Fukuda A, Wickman LT, Venkatareddy MP,et al.Urine podocin:nephrin mRNA ratio(PNR)as a podocyte stress biomarker[J].Nephrol Dial Transplant,2012,27(11):4079-4087.
[2] Li B,Hartono C,Ding R,et al.Noninvasive diagnosis of renal-allograftrejection by measurement of messenger RNA for perforin and granzyme B in urine[J].N Eng J Med,2001,344(13):947-954.
[3] Wang G,Lai FM,Lai KB,et al.Urinary messenger RNA expression of podocyte-associated molecules in patients with diabetic nephropathy treated by angiotensin-converting enzyme inhibitor and angiotensin receptor blocker[J].Eur JEndocrinol,2008,158(3):317-322.
[4] Matignon M,Ding R,Dadhania DM,et al.Urinary cell mRNA profiles and differential diagnosis of acute kidney graft dysfunction[J].J Am Soc Nephrol,2014,25(7):1586-1597.
[5] Jim B,Santos J,Spath F,et al.Biomarkers of diabetic nephropathy, thepresent and the future[J].Curr Diabetes Rev,2012,8(5):317-328.
[6] Kufer P,Zippelius A,Lutterbuse R,et al.Heterogeneous expression of MAGE-A genes in occult disseminated tumor cells:anovel multimarker reverse transcription-polymerase chain reaction for diagnosis of micrmetastaic disease[J].Cancer Res,2002,62(1):251-261.
[7] 王震炜,徐晓萍,徐 磊,等.血Cystatin C与尿NAG在急性肾功能损伤中的诊断应用[J].现代检验医学杂志,2011,26(2):125-126,128. Wang ZW,Xu XP,Xu L,et al.Serum Cystatin C and urinary NAG measurements for diagnosis of acute kidney injury[J].Journal of Modern Laboratory Medicine,2011,26(2):125-126,128.
[8] Szeto CC,Tam LS,Kwan BC,et al.Monitoring of urinary messenger RNA levels for the prediction of flare in systemic lupus erythematosus[J].Clin ChimActa,2012,413(3/4):448-455.

相似文献/References:

[1]肖传宇,王 宁,王小艳,等.血清胰岛素与慢性肾脏病的相关研究及其应用价值[J].现代检验医学杂志,2017,32(01):109.[doi:10.3969/j.issn.1671-7414.2017.01.030]
 XIAO Chuan-yu,WANG Ni,WANG Xiao-yan,et al.Correlation of Serum Insulin and Chronic Kidney Disease and Its Application Value[J].Journal of Modern Laboratory Medicine,2017,32(04):109.[doi:10.3969/j.issn.1671-7414.2017.01.030]
[2]卢美红,杨春兰,景蓉蓉,等.血清人附睾蛋白4表达与慢性肾脏病的相关性研究[J].现代检验医学杂志,2017,32(02):75.[doi:10.3969/j.issn.1671-7414.2017.02.020]
 LU Mei-hong,YANG Chun-lan,JING Rong-rong,et al.Study on Association with Expression in Patients with Chronic Kidney Disease[J].Journal of Modern Laboratory Medicine,2017,32(04):75.[doi:10.3969/j.issn.1671-7414.2017.02.020]
[3]杨 雄,尹爱萍.慢性肾脏病不同分期患者血清多项骨代谢指标变化与成纤维细胞生长因子23的相关性分析[J].现代检验医学杂志,2019,34(04):58.[doi:10.3969/j.issn.1671-7414.2019.04.014]
 YANG Xiong.,YIN Ai-ping.Analysis of the Correlation between Serum Bone Metabolism and Fibroblast Growth Factor 23 in Patients with Chronic Kidney Disease at Different Stages[J].Journal of Modern Laboratory Medicine,2019,34(04):58.[doi:10.3969/j.issn.1671-7414.2019.04.014]
[4]杨 雄,尹爱萍.慢性肾脏病患者血液中FGF23,TNF-α,RANTES 和IL-12 检测水平的相关性研究[J].现代检验医学杂志,2020,35(01):25.[doi:10.3969/j.issn.1671-7414.2020.01.007]
 YANG Xiong,YIN Ai-ping.Correlation between FGF23,TNF-α, RANTES and IL-12 Levels in the Bloodof Patients with Chronic Kidney Disease[J].Journal of Modern Laboratory Medicine,2020,35(04):25.[doi:10.3969/j.issn.1671-7414.2020.01.007]
[5]曹 雷a,汪隆海a,周 梅a,等.慢性肾心综合征患者血清9项心衰标志物水平联合检测的实验诊断价值研究[J].现代检验医学杂志,2021,36(05):90.[doi:10.3969/j.issn.1671-7414.2021.05.020]
 CAO Leia,WANG Long-haia,ZHOU Meia,et al.Study on Experimental Diagnostic Value for Combined Detection of 9 HeartFailure Markers in Serum of Patients with Chronic Renal Heart Syndrome[J].Journal of Modern Laboratory Medicine,2021,36(04):90.[doi:10.3969/j.issn.1671-7414.2021.05.020]

备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(NO.81270781)。
作者简介:王洁敏(1989-),女,在读研究生,研究方向:慢性肾脏病。
通讯作者: 顾乐怡(1973-),男,肾脏内科医师,E-mail:guleyi@aliyun.com。
更新日期/Last Update: 2016-08-10