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Journal of Clinical Microbiology, January 2000, p. 94-98, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a TT Virus DNA Quantification System Using Real-Time Detection PCR

Takanobu Kato,1 Masashi Mizokami,1,* Motokazu Mukaide,2 Etsuro Orito,1 Tomoyoshi Ohno,1 Tatsunori Nakano,1 Yasuhito Tanaka,1 Hideaki Kato,1 Fuminaka Sugauchi,1 Ryuzo Ueda,1 Noboru Hirashima,3 Kazuhide Shimamatsu,4 Masayoshi Kage,4 and Masamichi Kojiro4

Second Department of Medicine, Nagoya City University Medical School,1 and Department of Gastroenterology, Chyukyo Hospital,3 Nagoya, Center for Molecular Biology and Cytogenetics, SRL, Inc., Tokyo,2 and Department of Pathology, Kurume University School of Medicine, Kurume,4 Japan

Received 13 July 1999/Returned for modification 31 August 1999/Accepted 8 October 1999

Although TT virus (TTV) was isolated from a cryptogenic posttransfusion hepatitis patient, its pathogenic role remains unclear. It has been reported that the majority of the healthy population is infected with TTV. To elucidate the differences between TTV infection in patients with liver diseases and TTV infection in the healthy population, a quantification system was developed. TTV DNA was quantified by a real-time detection PCR (RTD-PCR) assay on an ABI Prism 7700 sequence detector. With this system, TTV DNA was quantified in 78 hepatitis C virus (HCV)-infected patients (63 with elevated serum alanine aminotransferase [ALT] levels and 15 with normal ALT levels) and in 70 voluntary blood donors (BDs). The quantification range was 2.08 to 7.35 log copies/ml. The intra-assay and interassay coefficients of variation were 0.37 to 6.33% and 0.60 to 7.07%, respectively. The mean serum TTV DNA levels in the HCV-infected patients with both elevated and normal ALT levels and BDs were 3.69 ± 0.89, 3.45 ± 0.76, and 3.45 ± 0.67 log copies/ml, respectively. Comparison of the serum TTV DNA levels among the HCV-infected patients revealed that they were not related to the serum ALT and HCV core protein levels or to the histopathological score on liver biopsy. This study showed that (i) the RTD-PCR assay for the detection of TTV was accurate and had a high degree of sensitivity, (ii) the mean serum TTV DNA level was similar among HCV-infected patients, irrespective of their ALT level, and also among BDs, and (iii) a high serum TTV DNA level does not affect the serum ALT and HCV levels or liver damage in HCV-infected patients.

* Corresponding author. Mailing address: Second Department of Medicine, Nagoya City University Medical School, Kawasumi, Mizuho, Nagoya 467-8601, Japan. Phone: 81-52-853-8216. Fax: 81-52-852-0849. E-mail: mizokami{at}med.nagoya-cu.ac.jp.

Journal of Clinical Microbiology, January 2000, p. 94-98, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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