Jpn. J. Infect. Dis., 60 (1), 29-32, 2007
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Presence of Hepatitis C Virus (HCV)-RNA in Peripheral Blood Mononuclear Cells in HCV Serum Negative Patients during Interferon and Ribavirin Therapy
Danuta Januszkiewicz-Lewandowska1,2, Jacek Wysocki1, Monika Pernak3, Karina Nowicka 2, Mariola Zawada2, Jolanta Rembowska2, Krzysztof Lewandowski2, Przemyslaw Mankowski1 and Jerzy Nowak2*
1University of Medical Sciences, 2Institute of Human Genetics of Polish Academy of Sciences, 3Department of Medical Diagnostics, Poznan, Poland
(Received July 12, 2006. Accepted November 24, 2006)
*Corresponding author: Mailing address: Institute of Human Genetics Polish Academy of Sciences, 60-479 Poznan, Strzeszynska 32, Poland. Tel: +48-618221312, Fax: +48-618233235, E-mail: firstname.lastname@example.org
SUMMARY: Identification of hepatitis C virus (HCV)-RNA in blood serum is crucial for hepatitis C diagnosis and for appropriate treatment. Detection of HCV-RNA in blood serum is used for therapy monitoring of patients with hepatitis C. Despite HCV-RNA elimination from blood serum during treatment in some patients, HCV viremia appears again after the completion of therapy. The aim of this study was to assess HCV-RNA in peripheral blood mononuclear cells (PBMCs) of hepatitis C patients in relation to HCV-RNA and antibodies to HCV in the serum. The study involved 71 patients undergoing anti-viral therapy (interferon and ribavirin). RNA isolated from serum and PBMCs was examined for the presence of HCV-RNA by an RT-PCR technique using specific oligonucleotide primers or by commercially available kits. In order to show the possible presence of HCV sequences in PBMCs, molecular DNA probes were constructed with a PCR amplicon and biotin-labelled by nick translation, and FISH and extended chromatin fibers in situ hybridization (ECFs-FISH) techniques were used. A 24-month follow-up study revealed that 34 out of 59 patients (58%) eliminated HCV-RNA from their sera. In the serum negative group, HCV-RNA was detected in PBMCs of 2 patients. The presence of HCV-RNA in PBMCs was confirmed by the FISH technique. In the ECFs-FISH procedure, no signal was found in all examined patients. Our data suggest that PBMCs infected with HCV can serve as a virus reservoir. HCV-RNA serum negative patients who have HCV-RNA in their leukocytes after completion of anti-viral therapy would be at great risk of hepatitis C recurrence. These HCV-RNA serum negative but PBMCs positive patients would be a potential source of HCV spread.
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