Practical experience of high throughput real time PCR in the routine diagnostic virology setting

被引:72
作者
Gunson, R. N. [1 ]
Collins, T. C. [1 ]
Carman, W. F. [1 ]
机构
[1] Gartnavel Royal Hosp, UK Clin Virol Network, W Scotland Specialist Virol Ctr, Glasgow G12 0YN, Lanark, Scotland
关键词
RT-PCR; virus diagnosis; sensitivity;
D O I
10.1016/j.jcv.2005.12.006
中图分类号
Q93 [微生物学];
学科分类号
071005 [微生物学]; 100705 [微生物与生化药学];
摘要
The advent of PCR has transformed the utility of the virus diagnostic laboratory. In comparison to traditional gel based PCR assays, real time PCR offers increased sensitivity and specificity in a rapid format. Over the past 4 years, we have introduced a number of qualitative and quantitative real time PCR assays into our routine testing service. During this period, we have gained substantial experience relating to the development and implementation of real-time assays. Furthermore, we have developed strategies that have allowed us to increase our sample throughput while maintaining or even reducing turn around times. The issues resulting from this experience (some of it bad) are discussed in detail with the aim of informing laboratories that are only just beginning to investigate the potential of this technology. (C) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:355 / 367
页数:13
相关论文
共 14 条
[1]
Basic principles of real-time quantitative PCR [J].
Arya, M ;
Shergill, IS ;
Williamson, M ;
Gommersall, L ;
Arya, N ;
Patel, HRH .
EXPERT REVIEW OF MOLECULAR DIAGNOSTICS, 2005, 5 (02) :209-219
[2]
Aslanzadeh J, 2004, ANN CLIN LAB SCI, V34, P389
[3]
Bustin Stephen A, 2004, J Biomol Tech, V15, P155
[4]
DAVIES AMC, 2003, SPECTROSC EUR, V15, P24
[5]
Draganov P, 2004, Akush Ginekol (Sofiia), V43, P38
[6]
Optimisation of PCR reactions using primer chessboarding [J].
Gunson, R ;
Gillespie, G ;
Carman, WF .
JOURNAL OF CLINICAL VIROLOGY, 2003, 26 (03) :369-373
[7]
Real-time RT-PCR detection of 12 respiratory viral infections in four triplex reactions [J].
Gunson, RN ;
Collins, TC ;
Carman, WF .
JOURNAL OF CLINICAL VIROLOGY, 2005, 33 (04) :341-344
[8]
Evaluation of a multiplex real-time reverse transcriptase PCR assay for detection and differentiation of influenza viruses A and B during the 2001-2002 influenza season in Israel [J].
Hindiyeh, M ;
Levy, V ;
Azar, R ;
Varsano, N ;
Regev, L ;
Shalev, Y ;
Grossman, Z ;
Mendelson, E .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (02) :589-595
[9]
ALLELIC DISCRIMINATION BY NICK-TRANSLATION PCR WITH FLUOROGENIC PROBES [J].
LEE, LG ;
CONNELL, CR ;
BLOCH, W .
NUCLEIC ACIDS RESEARCH, 1993, 21 (16) :3761-3766
[10]
Real-time PCR in the microbiology laboratory [J].
Mackay, IM .
CLINICAL MICROBIOLOGY AND INFECTION, 2004, 10 (03) :190-212