A polymerase chain reaction to detect a spliced late transcript of human cytomegalovirus in the blood of bone marrow transplant recipients.
A reverse transcription (RT) nested polymerase chain reaction (PCR) procedure is described for detecting RNA to a spliced late gene (SLG) of human cytomegalovirus (CMV), the product of which (175 bp) is easily differentiated in agarose gels from the product when the target is unspliced viral RNA or DNA (258 bp). The SLG-RT-PCR has been compared against a semi-quantitative PCR for CMV DNA in buffy-coat specimens collected weekly after bone marrow transplantation from 3 patients and against the results of culturing these specimens for CMV both by conventional virus isolation, based on the detection of cytopathic effect, and by the early detection of infected cells by staining with virus-specific monoclonal antibodies. The detection of CMV RNA by SLG-RT-PCR correlated well with the detection of infective virus but only when the results of both culture methods were combined, in that neither culture method alone was as sensitive as the SLG-RT-PCR. The presence of SLG RNA in the circulation is of value as a marker of active CMV infection.
Nelson
P N
PN
Department of Medical Microbiology, University of London, UK.
Rawal
B K
BK
Boriskin
Y S
YS
Mathers
K E
KE
Powles
R L
RL
Steel
H M
HM
Tryhorn
Y S
YS
Butcher
P D
PD
Booth
J C
JC
D001483:Base Sequence; D016026:Bone Marrow Transplantation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000097:blood / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D007962:Leukocytes / Q000821:virology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000097:blood; D012367:RNA, Viral / Q000097:blood*
D004279:DNA, Viral; D012333:RNA, Messenger; D012367:RNA, Viral