title: Comparison of cytomegalovirus viral load measure by real-time PCR with pp65 antigenemia for the diagnosis of cytomegalovirus disease in solid organ transplant patients.
abstract: Cytomegalovirus (CMV) infection is the most frequent complication in solid organ transplant recipients. Currently, the antigenemia assay is widely used to detect this infection, although its success is being questioned to a great extent nowadays. The aim of our study is to compare a quantitative real time PCR to measure CMV DNA to the antigenemia assay, for the diagnosis to CMV disease. For our research, we prospectively processed 1198 samples (plasma and peripheral blood leukocytes [PBMC]), which belonged to 158 transplant recipients. In every sample the detection of the pp65 antigen in PBMC was carried out, as well as the quantification of CMV DNA by PCR (Light Cycler, LC-PCR). For this process, FRET probes, which detect a 254-bp fragment from the CMV gB gene, were used. The dynamic range of the LC-PCR was 500 to 5.10(7) copies/mL plasma and from 62 to 6.10(6) copies/10(6) PBMC. Twenty-three episodes of cytomegalovirus (CMV) disease occurred in 22 out of 158 patients and PCR displayed levels of sensitivity and specificity of 100% and 67%, respectively. The antigenemia assay obtained values of 91% and 57%. We established a cutoff value of 10(3) copies/mL plasma and 315 copies/10(6) cells. According to these cutoff values, PCR showed levels of sensitivity, specificity, VPN and VPP of 95.6%, 81.6%, 99%, and 53% respectively. Moreover, the LC-PCR assay anticipated the antigenemia assay in 10 patients out of 22 who developed CMV disease and the appearance of any clinical symptoms in 12 out of 22 patients. In conclusion, we believe that the quantification of CMV DNA by LC-PCR is a superior assay to pp65 antigenemia test regarding the early diagnosis of CMV disease in solid organ transplant recipients.
authors.affiliation: Servicio de Microbiología, Hospital 12 de Octubre, Madrid, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Hernando
authors.forename: L
authors.initials: L
authors.lastname: Folgueira
authors.forename: C
authors.initials: C
authors.lastname: Lumbreras
authors.forename: R
authors.initials: R
authors.lastname: San Juan
authors.forename: S
authors.initials: S
authors.lastname: Maldonado
authors.forename: C
authors.initials: C
authors.lastname: Prieto
authors.forename: M J
authors.initials: MJ
authors.lastname: Babiano
authors.forename: J
authors.initials: J
authors.lastname: Delgado
authors.forename: A
authors.initials: A
authors.lastname: Andres
authors.forename: E
authors.initials: E
authors.lastname: Moreno
authors.forename: J M
authors.initials: JM
authors.lastname: Aguado
authors.forename: J R
authors.initials: JR
authors.lastname: Otero
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D006801:Humans; D016377:Organ Transplantation / Q000009:adverse effects*; D010750:Phosphoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011183:Postoperative Complications / Q000821:virology; D012367:RNA, Viral / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D012189:Retrospective Studies; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000235:genetics*
chemical_list: D010750:Phosphoproteins; D012367:RNA, Viral; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa