[Direct detection of human cytomegalovirus in urine specimens from bone marrow transplant patients by polymerase chain reaction].
Human cytomegalovirus (HCMV) infection is a major causative life-threatening agent that results in opportunistic infections in bone marrow transplant patients. Since antiviral therapy is available for severe HCMV infections, methods to rapidly identify infected patients are needed so that therapy can be promptly instituted. In this study, we used direct method to detect HCMV in urine specimens by nested polymerase chain reaction (nested-PCR) to monitor urinary excretion of HCMV in patients undergoing bone marrow transplantation. The viral DNA was amplified directly from preheated urine without further treatment prior to amplification. Five microliter of urine proved to give the most efficient amplification. The detection limit of the PCR assay for detection of HCMV was 10 copies/microl. The positive rate of the PCR assay and the tissue culture method for detection of HCMV were 15 of 61 (24.6%) and 4 of 61 (6.6%), respectively. Sensitivity and specificity of the PCR assay were 100% and 81% respectively. Based on these data, urinary excretion of HCMV in 21 patients undergoing bone marrow transplantation were monitored weekly by the PCR assay. As a result, in addition to a higher sensitivity, the PCR assay allowed to identify HCMV infection 3.1 (1 approximately 5) weeks earlier than culture method or antibody elevation. Repeated monitoring of virus excretion by this rapid and simple method was useful to promptly detect HCMV infection, allowing proper institution of antiviral drug therapy in patients undergoing bone marrow transplantation.
Ohshima
T
T
Central Clinical Laboratory, Tokai University Hospital, Isehara, Japan.
Miyachi
H
H
Fusegawa
H
H
Tanaka
S
S
Yabe
H
H
Yabe
M
M
Kato
S
S
Ando
Y
Y
D000293:Adolescent; D000328:Adult; D016026:Bone Marrow Transplantation; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D006801:Humans; D016867:Immunocompromised Host; D007223:Infant; D008297:Male; D016133:Polymerase Chain Reaction; D014556:Urine / Q000821:virology