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Clinical Chemistry 48: 1551-1559, 2002;
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(Clinical Chemistry. 2002;48:1551-1559.)
© 2002 American Association for Clinical Chemistry, Inc.

Quantitative Analysis of p40/p46 and p69/p71 Forms of 2',5'-Oligoadenylate Synthetase mRNA by Competitive PCR and Its Clinical Application

Akira Takahashi1, Yoshiaki Iwasaki3a, Jiro Miyaike1, Hideaki Taniguchi1, Hiroyuki Shimomura3, Tadashi Hanafusa4, Yasuhiro Yumoto4, Akio Moriya1, Norio Koide2 and Takao Tsuji1

Departments of
1 Medicine and Medical Science and
2 Laboratory Medicine, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
3 First Department of Internal Medicine, Okayama University Medical School, Okayama 700-8558, Japan.

4 Radioisotope Center, Okayama University, Okayama 700-8558, Japan.

aAddress correspondence to this author at: First Department of Internal Medicine, Okayama University Medical School, 2-5-1, Shikata-cho, Okayama 700-8558, Japan. Fax 81-86-225-5991; e-mail yiwasaki{at}cc.okayama-u.ac.jp.

Background: 2',5'-Oligoadenylate synthetases (2-5AS) are type I interferon (IFN)-induced proteins with antiviral capacity. Three major forms of 2-5AS with distinct enzymatic activities have been described in IFN-treated human cells. We measured distinct forms of 2-5AS mRNA to analyze the relationship with its enzymatic activity and response to IFN therapy in chronic hepatitis C.

Methods: We established a method to quantify p40/p46 and p69/p71 forms of 2-5AS mRNA by use of reverse transcription followed by competitive PCR. The 2-5AS mRNA concentrations were measured in peripheral blood mononuclear cells from 40 patients with chronic hepatitis C and 28 control individuals.

Results: Reconstitution experiments and comparison with Northern blot analyses revealed that our method accurately and linearly quantified 2-5AS mRNA. 2-5AS mRNA concentrations and 2-5AS enzymatic activity were correlated (P <0.03). Our data demonstrated a correlation in 2-5AS mRNA between p40/p46 and p69/p71 (P <0.02), indicating a similar regulation of the expression of these genes. Our data also demonstrated that pretreatment concentrations of 2-5AS mRNA correlated with responses to IFN therapy in chronic hepatitis C.

Conclusions: Our method for measuring 2-5AS mRNA concentrations could provide an important marker for selecting patients for IFN therapy and may be useful for the development of more effective therapeutic strategies for chronic hepatitis C.






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