title: Early detection of cytomegalovirus (CMV) infection in bone marrow transplant patients by reverse transcription-PCR for CMV spliced late gene UL21.5: a two site evaluation.
abstract: BACKGROUND
  Bone marrow transplant (BMT) patients at risk of developing cytomegalovirus (CMV) pneumonitis are identified routinely by the early detection of virus in blood. For early diagnosis of CMV infection, the RNA-based approach demonstrates advantages when compared with the current CMV antigen and DNA detection methods.
  
  
  OBJECTIVE
  We have evaluated our previously developed reverse transcription-polymerase chain reaction (RT-PCR) to a spliced late CMV gene (SLG; J. Virol. Methods 56 (1996), 139) to monitor CMV infection in BMT patients at two clinical sites. The diagnostic value of the SLG RT-PCR was compared with the routine CMV antigen and DNA detection methods.
  
  
  METHODS
  Weekly blood samples from BMT patients were tested for CMV during the first 3 months post-transplant. The qualitative SLG RT-PCR, semiquantitative DNA PCR, and viral antigen tests were compared. The RNA and DNA PCR results were analysed in terms of their temporal relationship and consistency of CMV detection and compared with CMV infection diagnosed by viral antigen tests.
  
  
  RESULTS
  Of the 101 BMT recipients studied, 25 developed CMV antigenemia and/or DNAemia resulting in symptomatic infection in two patients. All CMV PCR-positive patients were either CMV seropositive pretransplant or received marrow from seropositive donor. The highest incidence of CMV infection was seen in seropositive recipients (R+) irrespective of the donor's status. Detection of CMV infection by SLG RNA preceded CMV DNA detection by 0-2 weeks (median 1 week) and CMV antigen detection by 0-8 weeks (median 3 weeks). Once detected, the SLG RNA remained consistently positive before antiviral treatment was commenced. Both the SLG RNA and CMV DNA detection methods had the same clinical sensitivity, specificity, positive and negative predictive values of 100, 94, 80 and 100%, respectively.
  
  
  CONCLUSIONS
  The RT-PCR for SLG RNA proved to be the earliest indicator of CMV infection in BMT patients demonstrating a sustained pattern of CMV detection during the 3 months post-transplant period. Although very similar in its diagnostic performance to CMV DNA PCR the SLG RNA RT-PCR does not require quantitation and provides an efficient and ongoing indication of active CMV infection.
authors.affiliation: Department of Medical Microbiology, St. George's Hospital Medical School, Cranmer Terrace, London SW17 ORE, UK. yboriski@sghms.ac.uk
authors.forename: Yu S
authors.initials: YS
authors.lastname: Boriskin
authors.forename: K
authors.initials: K
authors.lastname: Fuller
authors.forename: R L
authors.initials: RL
authors.lastname: Powles
authors.forename: I B
authors.initials: IB
authors.lastname: Vipond
authors.forename: P S
authors.initials: PS
authors.lastname: Rice
authors.forename: J C
authors.initials: JC
authors.lastname: Booth
authors.forename: E O
authors.initials: EO
authors.lastname: Caul
authors.forename: P D
authors.initials: PD
authors.lastname: Butcher
mesh_terms: D000293:Adolescent; D000956:Antigens, Viral / Q000032:analysis; D016026:Bone Marrow Transplantation; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000209:etiology*; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D011183:Postoperative Complications; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral; D012367:RNA, Viral