Primer-mediated enzymatic amplification of cytomegalovirus (CMV) DNA. Application to the early diagnosis of CMV infection in marrow transplant recipients.
A nucleic acid amplification procedure, the polymerase chain reaction (PCR), has been used to establish a diagnostic assay for the identification of cytomegalovirus (CMV) immediate-early sequences in clinical specimens. Preliminary testing against virus-infected cell cultures indicated that the PCR assay was highly CMV-specific, recognizing both wild-type and laboratory strains of CMV. There was no cross-reactivity with human DNA or with DNA from other herpes viruses. The sensitivity of the assay, using cloned CMV AD169 Eco RI fragment-J as template, was 1 viral genome per 40,000 cells. In a prospective study of CMV infection in bone marrow transplant recipients, the PCR assay correctly identified four patients with confirmed CMV infection. In three of these patients who were followed longitudinally, correlation of DNA reactivity with CMV culture and CMV antibody status over time indicated that DNA was the most sensitive marker for the diagnosis of CMV infection.
Cassol
S A
SA
Canadian Red Cross, Blood Transfusion Service, Calgary, Alberta.
Poon
M C
MC
Pal
R
R
Naylor
M J
MJ
Culver-James
J
J
Bowen
T J
TJ
Russell
J A
JA
Krawetz
S A
SA
Pon
R T
RT
Hoar
D I
DI
D000293:Adolescent; D000328:Adult; D001782:Blood Donors; D016026:Bone Marrow Transplantation; D003587:Cytomegalovirus / Q000235:genetics* / Q000254:growth & development; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000235:genetics / Q000382:microbiology; D004279:DNA, Viral / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase; D005260:Female; D005784:Gene Amplification; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D011658:Pulmonary Fibrosis / Q000209:etiology; D019914:Taq Polymerase
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase