title: Comparison of three PCR techniques for detecting cytomegalovirus (CMV) DNA in serum, detection of early antigen fluorescent foci and culture for the diagnosis of CMV infection.
abstract: Three PCR assays were developed for detection of cytomegalovirus (CMV) DNA in serum and were evaluated with samples from organ transplant recipients. The Qiamp Blood Kit was efficient for extraction of DNA from sera. Single-round PCR of a 293-bp region of CMV DNA was sensitive and highly specific for CMV targets and was more sensitive than detection of early antigen fluorescent foci (DEAFF) testing or isolation of CMV from buffy coat by cell culture. The results of a significant proportion of buffy coat samples were not interpretable because of toxicity in conventional culture or DEAFF tests. A non-competitive quantitative PCR test and semi-quantitative PCR test for the detection of CMV DNA in serum yielded comparable results for samples taken serially from three bone marrow transplant recipients. Single-round PCR was superior to conventional techniques for the diagnosis of CMV infection, was simple to perform and was completed rapidly. The semi-quantitative technique has added advantages where quantification is important.
authors.affiliation: University of Cambridge School of Clinical Medicine, Hills Road, Cambridge CB2 2QQ.
authors.forename: P C
authors.initials: PC
authors.lastname: Evans
authors.affiliation: Clinical Microbiology and Public Health Laboratory, Hills Road, Cambridge CB2 2QQ.
authors.forename: J J
authors.initials: JJ
authors.lastname: Gray
authors.affiliation: Clinical Microbiology and Public Health Laboratory, Hills Road, Cambridge CB2 2QQ.
authors.forename: T G
authors.initials: TG
authors.lastname: Wreghitt
authors.affiliation: Department of Haematology, Addenbrooke's NHS Trust, Hills Road, Cambridge CB2 2QQ.
authors.forename: R E
authors.initials: RE
authors.lastname: Marcus
authors.affiliation: University of Cambridge School of Clinical Medicine, Hills Road, Cambridge CB2 2QQ.
authors.forename: G J M
authors.initials: GJM
authors.lastname: Alexander
mesh_terms: D000328:Adult; D000956:Antigens, Viral / Q000032:analysis*; D016026:Bone Marrow Transplantation; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D017874:Immediate-Early Proteins / Q000032:analysis*; D016031:Liver Transplantation; D008297:Male; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D012044:Regression Analysis; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral; D017874:Immediate-Early Proteins; C044582:immediate-early proteins, cytomegalovirus