title: Comparison of nested PCR for detection of DNA in plasma with pp65 leukocytic antigenemia procedure for diagnosis of human cytomegalovirus infection.
abstract: A nested PCR was used for the detection of human cytomegalovirus (HCMV) DNA in plasma. The presence of HCMV DNA and its correlation to pp65 leukocytic antigenemia were investigated with 299 blood samples from 45 organ transplant recipients and 63 AIDS patients. Of the 53 samples positive by nested PCR, 52 (98%) were also positive for leukocytic antigenemia and 23 had high levels of antigenemia (> 50 positive cells per 2 x 10(5) leukocytes). Of the 246 samples negative in PCR, only 3 (1.2%) had highly positive antigenemia. For 15 patients having a high antigenemia level in the course of their disease, consecutive blood samples were studied and also assessed for viremia in culture. The extent to which HCMV DNA, detected by PCR, was present in plasma correlated with increased levels of HCMV leukocytic antigenemia for six of the eight AIDS patients and for all the organ transplant recipients. Positivity for HCMV DNA in PCR and for viremia in cell culture was usually restricted to the highest antigenemia levels. From a total of 69 blood samples, PCR and culture gave positive results, respectively, for 17 of 32 samples (53%) and 14 of 32 samples (43%) from transplant recipients and for 15 of 37 samples (40%) and 9 of 37 samples (24%) from AIDS patients. Our findings have shown a strong correlation between high levels of leukocytic antigenemia and HCMV DNA in plasma. The detection of HCMV DNA in plasma by this nested PCR can prove HCMV dissemination in blood, but it lacks the rapidity and simplicity of the leukocytic pp65 antigenemia procedure.
authors.affiliation: Virus Laboratory, University Hospital Caen, France.
authors.forename: F
authors.initials: F
authors.lastname: Freymuth
authors.forename: E
authors.initials: E
authors.lastname: Gennetay
authors.forename: J
authors.initials: J
authors.lastname: Petitjean
authors.forename: G
authors.initials: G
authors.lastname: Eugene
authors.forename: B
authors.initials: B
authors.lastname: Hurault de Ligny
authors.forename: J P
authors.initials: JP
authors.lastname: Ryckelynck
authors.forename: C
authors.initials: C
authors.lastname: Legoff
authors.forename: P
authors.initials: P
authors.lastname: Hazera
authors.forename: C
authors.initials: C
authors.lastname: Bazin
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000382:microbiology; D001483:Base Sequence; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000382:microbiology*; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood*; D006801:Humans; D007962:Leukocytes / Q000382:microbiology*; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000097:blood*; D016133:Polymerase Chain Reaction; D011183:Postoperative Complications / Q000382:microbiology; D014180:Transplantation; D014763:Viral Matrix Proteins / Q000097:blood*; D014766:Viremia / Q000382:microbiology*
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa