Development of fluorescent quantitation PCR method for detection of urinary CK20

GUO Bin; et al

您当前的位置：

首页 >

文章列表页 >

Development of fluorescent quantitation PCR method for detection of urinary CK20

更新时间：2024-01-17

- Development of fluorescent quantitation PCR method for detection of urinary CK20
- Journal of Chongqing Medical University Vol. 34, Issue 6, Pages: 752-755(2009)
- 作者机构：
  
  1. 重庆医科大学医学检验系临床检验诊断学教育部重点实验室
  2. 重庆医科大学附属第一医院泌尿外科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R737.14
- Published：2009，
扫描看全文
[1]郭斌,罗春丽,荀春华,谢建红,吴小候,蒲军.实时荧光定量PCR检测尿液CK20方法的建立[J].重庆医科大学学报,2009,34(06):752-755.

GUO Bin, et al. Development of fluorescent quantitation PCR method for detection of urinary CK20. [J]. Journal of Chongqing Medical University 34(6):752-755(2009)
[1]郭斌,罗春丽,荀春华,谢建红,吴小候,蒲军.实时荧光定量PCR检测尿液CK20方法的建立[J].重庆医科大学学报,2009,34(06):752-755. DOI：

GUO Bin, et al. Development of fluorescent quantitation PCR method for detection of urinary CK20. [J]. Journal of Chongqing Medical University 34(6):752-755(2009) DOI：

摘要

目的:建立一种简便、快速、灵敏度和特异性较好的定量检测尿液CK20方法

用于膀胱移行细胞癌（Transitional cell car-cinoma of the bladder

TCCB）早期诊断、预后及复发的评估。方法:培养膀胱癌T24细胞

针对CK20基因保守序列设计合成两对引物和Taqman探针

将PCR扩增产物片段克隆

作为定量检测的标准品

优化反应条件

建立快速检测CK20mRNA的实时荧光定量PCR方法

并进行方法学评价。结果:荧光定量PCR检测尿液CK20方法能较好的定量检出尿液CK20mRNA

检测范围为102～109copies/μl

灵敏度为102copies/μl

特异性好

批内、天间重复性（Coefficientofvariation

CV）分别为0.89%和2.45%。结论:成功建立了快速检测尿液CK20的荧光定量PCR方法

为进一步开发出用于TCCB早期诊断、预后及复发评估的定量检测试剂盒奠定了基础。

Abstract

Objective:To develop a novel real-time fluorescence PCR method using Taqman probe for convenient

fast

sensitive and specific detection of urinary CK20 and use in early diagnosis and follow-up of transitional cell carcinoma of the bladder(TCCB).Methods:CK20 gene from cultured T24 cells was amplified by conventional RT-PCR

the standard quantitative plasmid was constructed by T-Aclone method.Taqman probe and primers was designed according to the sequence of CK20 cloned gene

the real-time PCR method for determination of CK20 mRNA was established and evaluated.Results:The developed real-time PCR method showed high sensitivity(10 2copies /μl) and good specificity;the linear range was 102 ~109 copies /μl

the coefficient variation(CV) was 0.89 % in intra-assay and 2.45% in day to day.Conclusion:A novel real-time fluorescence quantitative PCR method for detection of CK20 was established.It provided a basis for developing new diagnostic kits using the Taqman probe for clinical application.

关键词

膀胱移行细胞癌CK20实时荧光定量PCR

Keywords

Transitional cell carcinoma of the bladderCK20real-time PCR

references

Views

102

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Methods of DNA extraction from bacteria for PCR and Real-time PCR

Expressions and significances of promoting apoptosis gene programmed cell death 5 in non-small cell lung cancer

Y chromosome micro-deletions in male infertility screened by real-time PCR

Related Author

HU Xiao-hong

et al

刘昕

黄正根

袁科

ZHAO Qing

ZHANG Mingchuan

MEI Tonghua

Related Institution

Department of Cell Biology and Genetics,College of Basic Medicine,Chongqing Medical University

第三军医大学分子遗传学教研室

Department of Respiratory Medicine，the First Affiliated Hospital，Nanchang University

Department of Respiratory Medicine，Tongliang People’s Hospital of Chongqing

Center for Clinical Molecular Medical Detection，The First Affiliated Hospital of Chongqing Medical University

⁰