Rapid quantitative analysis of human cytomegalovirus DNA by the real-time polymerase chain reaction method.
BACKGROUND
Human cytomegalovirus (CMV) infection is a progressive and life-threatening complication in immunocompromised patients even now. Therefore, early and accurate treatment based on rapid and certain detection is needed to prevent fatal CMV infection diseases.


OBJECTIVE
To study a quicker, simpler, and less expensive method of quantitative analysis using real-time polymerase chain reaction based on the SYBR Green I method of CMV detection for appropriate treatment of CMV infection in immunocompromised patients.


METHODS
We quantified 50 samples tested by direct immunoperoxidase staining of leukocytes with peroxidase-labeled monoclonal antibody (C7-HRP test), 30 samples from healthy persons, and 47 samples from 7 patients suspected of having CMV infection diseases. We used the primer set in the pp65 gene of CMV and whole blood without a preparatory process. The setting for the study was the First Department of Pathology, Kurume University School of Medicine, St Mary's Hospital, and the Gene Section of the Clinical Laboratory at St Mary's Hospital, Fukuoka, Japan.


RESULTS
The results obtained with this method corresponded well with conventional C7-HRP tests and demonstrated excellent reproduction. Additionally, the results were better correlated with the clinical course than were C7-HRP tests.


CONCLUSIONS
This method was more useful than the C7-HRP test as a rapid diagnostic test for early treatment of CMV infection. This test also demonstrated its usefulness for monitoring CMV infection during treatment using ganciclovir. Moreover, it was quicker, simpler, and cheaper than other real-time polymerase chain reaction methods.
Numata
Sanae
S
First Department of Pathology, Kurume University School of Medicine, Kurume-shi, Fukuoka, Japan. s-numa@st-mary-med.or.jp <s-numa@st-mary-med.or.jp>
Nakamura
Yasuhiro
Y
Imamura
Yutaka
Y
Honda
Junichi
J
Momosaki
Seiya
S
Kojiro
Masamichi
M
D000328:Adult; D000368:Aged; D000741:Anemia, Aplastic / Q000150:complications / Q000821:virology; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000914:Antibodies, Viral / Q000378:metabolism; D003199:Computer Systems; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000150:complications / Q000175:diagnosis / Q000188:drug therapy; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D015774:Ganciclovir / Q000627:therapeutic use; D006801:Humans; D016867:Immunocompromised Host / Q000235:genetics; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000150:complications / Q000821:virology; D015470:Leukemia, Myeloid, Acute / Q000150:complications / Q000821:virology; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000150:complications / Q000821:virology; D008297:Male; D008875:Middle Aged; D010750:Phosphoproteins / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D014763:Viral Matrix Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017931:DNA Primers; D004279:DNA, Viral; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa; D015774:Ganciclovir