Normalized quantification of human cytomegalovirus DNA by competitive real-time PCR on the LightCycler instrument.

ABSTRACT: The development of a novel normalized quantitative competitive real-time PCR on the LightCycler instrument (NQC-LC-PCR) and its application to the quantification of cytomegalovirus (CMV) DNA in clinical samples are described. A heterologous competitor DNA was spiked into test samples and served as an internal amplification control. The internal control (IC) DNA in the test samples was coamplified with the CMV DNA and was tested against a calibrator sample that contained equal amounts of IC DNA and CMV reference standard DNA. An algorithm was developed to normalize possible varying amplification efficiencies between the standard and the samples. After normalization, CMV DNA copy numbers were determined in absolute terms. In a routine clinical setting, normalized quantification by NQC-LC-PCR

SUBMITTER: Stocher M 

PROVIDER: S-EPMC154596 | biostudies-literature | 2002 Dec

REPOSITORIES: biostudies-literature