nav emailalert searchbtn searchbox tablepage yinyongbenwen piczone journalimg journalInfo journalinfonormal searchdiv searchzone qikanlogo popupnotification paper paperNew
2005, 12, 97-101
实时荧光定量PCR技术
基金项目(Foundation): 国家科技部猪瘟监测技术平台(2004BA519A19-加强); 国家“十五”攻关子课题(2004BA514A16-1)
邮箱(Email):
DOI: 10.16437/j.cnki.1007-5038.2005.12.025
摘要:

荧光定量PCR技术是一种目前国内外应用较为广泛的定量PCR技术,是通过始点定量和荧光检测系统实时监测累积荧光强度而实现的,具有灵敏度高、特异性强、线性关系好、操作简单等优点,不但能够用于基因定量检测,还能准确定量疾病相关基因的表达,现已广泛应用于人类和动物疾病的快速检测、定量分析、早期诊断、基因分型等方面。它可以采用绝对定量和相对定量2种方式实现定量检测,但都有各自的优缺点,需根据具体的试验选择合适的定量方式,从而得到更准确的试验结果。提高荧光定量PCR技术的特异性和灵敏度需从多方面着手,主要指特异性探针及引物的设计,选择合适的Mg2+浓度、引物和探针浓度、循环数等。

Abstract:

KeyWords:
参考文献

[1]Magnus L,Charles H.Dynamic range and reproducibility ofhepatitis B virus(HBV)DNA detection and quantification bycobas Taqman HBV,a real-time semiautomated assay[J].JClin Microbiol,2005,43:4251-4254.

[2]张淑云,刘伟,谷鸿喜,等.实时荧光定量PCR检测血清中HCV-RNA[J].世界华人消化杂志,2004,(12)6:1464-1465.

[3]张冬雷,施健,崔之础,等.实时荧光定量检测HCV RNA含量的临床意义[J].世界华人消化杂志,2005,13(1):39-43.

[4]Nathalie D,Axelle D,Véronique S,et al.Quantification ofHuman immunodeficiency virus type 1 proviral load by a Taq-Man real-time PCR assay[J].J Clin Microbiol,2001,39:1303-1310.

[5]Robert M W,Stella L,Michael G,et al.Measurement of ep-stein-barr virus DNA loads in whole blood and plasma by Taq-Man PCR and in peripheral blood lymphocytes by competitivePCR[J].J Clin Microbiol,2003,41:5245-5249.

[6]冯燕,卢亦愚,严菊英,等.荧光定量RT-PCR技术快速检测SARS病毒核酸[J].中国病毒学,2005,20(3):228-231.

[7]Jones C D,Yeung C,Zehnder J L,et al.Comprehensive vali-dation of a real-time quantitative bcr-abl assay for clinical labo-ratory use[J].J Clin Pathol,2003,120(1):42-48.

[8]Ueno H,Yoshida K,Hirai T.Quantitative detection of carci-noembryonic antigen messenger RNA in the peritoneal cavityof gastric cancer patients by real-time quantitative reverse tran-scription polymerase chain reaction[J].Anticancer Res,2003,23(2C):1701-1708.

[9]Nieuwenhuis E J,Jaspars L H,Castelijns J A,et al.Quanti-tative molecular detection of minimal residual head and neckcancer in lymph node aspirates[J].Clin Cancer Res,2003,9(2):755-761.

[10]Reis P P,Rogatto S R,Kowalski L P,et al.Quantitative re-al-time PCR identifies a critical region of deletion on 22q13 re-lated to prognosis in oral cancer[J].Oncogene,2002,21(42):6480-6487.

[11]Summers K E,Davies A J,Matthews J,et al.The relativerole of peripheral blood and bone marrow for monitoring mo-lecular evidence of disease in follicular lymphoma by quantita-tive real-time polymerase chain reaction[J].Br J Haematol,2002,118(2):563-566.

[12]Mcgoldrick A,Lowings J P,Ibata G,et al.A novel approachto the detection of classical swine fever virus by RT-PCR witha fluorogenic probe(TaqMan)[J].Journal of VirologicalMethods,1998,72:125-135.

[13]陈玉栋,张楚瑜,邹俊煊,等.建立快速定量检测猪瘟兔化弱毒苗的荧光定量PCR技术[J].中国病毒学,2003,18(2):124-128.

[14]Letellier C,Kerkhofs P.Real-time PCR for simultaneous de-tection and genotyping of bovine viral diarrhea virus[J].Journal of Virological Methods,2003,114:21-27.

[15]Piet A,van Rijn,Gerard J W,et al.Detection of economical-ly important viruses in boar semen by quantitative RealTimePCRTM technology[J].Journal of Virological Methods,2004,120:151-160.

[16]张鹤晓,赖平安,刘环,等.荧光RT-PCR检测活禽和禽组织中禽流感病毒的研究[J].检验检疫科学,2004,4(1):1-5.

[17]朱文斯,赖平安,黄茜华,等.荧光RT-PCR快速检测禽流感病毒H5亚型的研究[J].中国医药导刊,2003,(1):7-30.

[18]花群义,徐自忠,杨云庆,等.TaqManR○RT-PCR对水疱性口炎病毒的鉴定检测[J].动物医学进展,2004,5(2):64-68.

[19]Bellin T,Pulz M,Matussek A,et al.Rapid detection of en-terohemorrhagic escherichia coli by real-time PCR with fluo-rescent hybridization probes[J].J Clin Microb,2001,39:370-374.

[20]Schalasta G,Eggers M,Schmid M,et al.Analysis of humancytomegalovirus DNA in urines of newborns and infants bymeans of a new ultrarapid real-time PCR-system[J].J ClinVirol,2000,19:175-185.

[21]张智俊,李亚玲,田兴军.用于SARS冠状病毒快速诊断的分子信标探针设计[J].生物信息学,2004,2(3):23-26.

[22]姜晓峰,王雅杰,张锟.毛细管电泳技术与分子信标技术结合检测基因[J].中国实验诊断学,2003,(3):185-189.

[23]汲振余,欧阳军,司源,等.乙型肝炎病毒Amplisensor定量分析[J].郑州大学学报(医学版),2004,9(6):1001-1004.

[24]廖锦良,谭耀驹,陈志成,等.Amplisensor-聚合酶反应定量检测脑脊液中结核分支杆菌DNA的研究[J].中国防痨杂志,2001,23(4):245-247.

[25]谭耀驹,李一耕,张言斌,等.AmpliSensor-聚合酶链反应定量检测肺结核患者外周血结核分支杆菌DNA[J].中华结核和呼吸杂志,1999,22(8):481-483.

[26]谭耀驹,李一耕,谭守勇,等.AmpliSensor-聚合酶链反应技术检测结核分支杆菌及临床应用[J].中华结核和呼吸杂志,1998,21(2):79-81.

[27]Wolfgang K,Jonathan N,Karas S,et al.Fluorogenic primersfor real-time quantitative PCR[J].American BiotechnologyLaboratory,2003,(7):52-56.

基本信息:

DOI:10.16437/j.cnki.1007-5038.2005.12.025

中图分类号:Q503

引用信息:

[1]蒋春燕,王泰健,王琴,等.实时荧光定量PCR技术[J].动物医学进展,2005(12):97-101.DOI:10.16437/j.cnki.1007-5038.2005.12.025.

基金信息:

国家科技部猪瘟监测技术平台(2004BA519A19-加强); 国家“十五”攻关子课题(2004BA514A16-1)

发布时间:

2005-12-20

出版时间:

2005-12-20

引用

GB/T 7714-2015 格式引文
MLA格式引文
APA格式引文
检 索 高级检索