In situ RT-PCR can distinguish between productive and latent cytomegalovirus infection in the blood cells of bone marrow transplant recipients.
Thirty-four peripheral blood leukocyte samples from bone marrow transplant (BMT) recipients were examined for Human cytomegalovirus (HCMV) phosphoprotein 65 (pp65), DNA and late transcripts. Twenty seven samples were positive for pp65 in the cytoplasm by immunofluorescent assay (IFA). Viral DNA was confirmed in 26 samples by nested PCR (nPCR). Using in situ RT-PCR, viral late transcripts were found in 19 samples, positive also by IFA and nPCR; these samples were considered indicative of productive viral infection. Five samples, positive by nPCR but negative by IFA and in situ RT-PCR, were considered to represent latent viral infection. In 8 samples, positive by IFA and nPCR but negative by in situ RT-PCR, apparently phagocytosis of viral particles took place.
Zawilinska
B
B
Department of Virology, Chair of Microbiology, Jagiellonian University Medical College, Cracow, Poland.
Bulek
K
K
Kopec
J
J
Daszkiewicz
E
E
Rojek-Zakrzewska
D
D
Kosz-Vnenchak
M
M
D016026:Bone Marrow Transplantation / Q000009:adverse effects*; D003587:Cytomegalovirus / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004279:DNA, Viral / Q000097:blood; D005455:Fluorescent Antibody Technique; D006801:Humans; D010750:Phosphoproteins / Q000097:blood*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics; D014763:Viral Matrix Proteins / Q000097:blood*; D017735:Virus Latency
D004279:DNA, Viral; D010750:Phosphoproteins; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; C040495:glycoprotein B, Simplexvirus