Development and evaluation of a real-time quantitative PCR for the detection of human cytomegalovirus

被引：50
|
作者：
Kearns, AM
Guiver, M
James, V
King, J
机构：
[1] Newcastle Gen Hosp, Publ Hlth Lab, Newcastle Upon Tyne NE4 6BE, Tyne & Wear, England
[2] Withington Hosp, Publ Hlth Lab, Manchester M20 2LR, Lancs, England
[3] Cent Publ Hlth Lab, London NW9 5HT, England
[4] Quality Assurance Lab, London NW9 5HT, England
[5] Publ Hlth Lab, Exeter EX2 5AD, Devon, England
来源：
JOURNAL OF VIROLOGICAL METHODS | 2001年 / 95卷 / 1-2期
关键词：
cytomegalovirus; viral load; real-time PCR; quantitative PCR; transplant patients;
D O I：
10.1016/S0166-0934(01)00307-X
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A novel real-time quantitative PCR (QPCR) assay is described for monitoring CMV DNA load in clinical specimens using the LightCycler (TM). The assay is rapid (< 40 min), easy to carry out, robust, reliable and is capable of detecting from 10 to over 2 x 10(5) CMV DNA copies with a wide linear range. Amplification and detection occur simultaneously. avoiding the need for post-PCR analysis and thereby minimising the risk of carryover contamination. The assay proved to be accurate, specific and reproducible when evaluated in three different laboratories. In addition. LightCycler (TM) results were comparable with those of TaqMan (TM). an independent real-time QPCR assay. (C) 2001 Elsevier Science B.V. All rights reserved.
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页码：121 / 131
页数：11
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