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Journal of Clinical Microbiology, September 2003, p. 4382-4387, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4382-4387.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Real-Time PCR Assay Compared to Nested PCR and Antigenemia Assays for Detecting Cytomegalovirus Reactivation in Adult T-Cell Leukemia-Lymphoma Patients

Junji Ikewaki,^* Eiichi Ohtsuka, Rie Kawano, Masao Ogata, Hiroshi Kikuchi, and Masaru Nasu

Department of Infectious Diseases, Oita Medical University, Oita, Japan

Received 25 March 2003/ Returned for modification 14 April 2003/ Accepted 14 June 2003

We analyzed the efficiency of the quantitative real-time PCR assay for cytomegalovirus (CMV) reactivation in adult T-cell leukemia-lymphoma (ATL) patients and compared the results with those obtained with qualitative nested PCR and antigenemia assays. The viral load obtained by the real-time PCR assay closely paralleled the number of antigen-positive cells obtained with the antigenemia assay. Real-time PCR revealed that a large number of DNA copies could be present even in samples assessed as negative or low in antigen-positive cells (0 to 10 antigen-positive cells/50,000 cells) by antigenemia assay. CMV copy numbers did not differ between the negative and low-antigen-positive groups. When the input concentration for real-time PCR assay was 2,500 to 5,000 copies/ml, the positivity rate for the nested PCR assay was 47.3%, while the positivity rate was more than 90% at an input concentration of 50,000 copies/ml. Real-time PCR is more sensitive than the antigenemia and nested PCR assays. Moreover, real-time PCR was able to detect CMV reactivation earlier than the antigenemia and nested PCR assays through the use of longitudinal analysis in four ATL patients with CMV pneumonia. In longitudinal assessments, analysis of the results suggested that a cutoff level of 5,000 copies/ml might be used to initiate treatment. Real-time PCR is more suitable for monitoring CMV reactivation in ATL patients than the antigenemia and nested PCR assays. CMV viral loads of 5,000 copies/ml are proposed as the cutoff for initiating antiviral therapy in ATL patients.

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* Corresponding author. Mailing address: Department of Infectious Disease, Oita Medical University, Hasama-Machi, Oita 879-5593, Japan. Phone: 81-97-586-5804. Fax: 81-97-549-4245. E-mail: ikewaki{at}oita-med.ac.jp.

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Journal of Clinical Microbiology, September 2003, p. 4382-4387, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4382-4387.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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