Development of a mass spectrometry-based quantitative assay for hepatitis virus detection

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K08937
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Ehime University
• Principal Investigator: Takemori Nobuaki 愛媛大学, 学術支援センター, 講師 (40533047)
• Co-Investigator(Kenkyū-buntansha): 鈴木 哲朗 浜松医科大学, 医学部, 教授 (00250184)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 質量分析 / 選択反応モニタリング / バイオマーカー / 肝炎ウイルス

Outline of Final Research Achievements

Serological diagnosis of hepatitis B virus is commonly performed with immunoassays against the HBV surface antigen (HBsAg). However, an alternative approach for HBsAg detection is currently required because of the emergence of immune-escape mutations, which cause false negative results in diagnostic tests. In this study, we developed an antibody-free strategy for HBsAg quantitation using targeted proteomics. By profiling tryptic HBsAg digests, we identified signature peptides that enabled the establishment of a reliable targeted assay for HBsAg quantitation, using selected reaction monitoring mass spectrometry. The established assay detected HBsAg at the sub-picogram level in a 100-nanoliter serum sample, and parallel monitoring of amino acid variations in signature peptides facilitated the determination of HBsAg genotypes. Our results demonstrate the feasibility of mass spectrometry-based diagnosis of HBV infection as an alternative method to conventional antibody-based diagnosis.

Free Research Field

分析化学

Academic Significance and Societal Importance of the Research Achievements

質量分析をベースとした検査法を新たに開発することで、従来のイムノアッセイ法で問題とされてきたエスケープ変異体による感染リスクの低減が可能となり、肝炎ウイルス感染の拡大防止に寄与できる。将来的には、新規ウイルスや、イムノアッセイの確立されていないウイルスに対する質量分析ベースの血液診断法を確立することにより感染症一般への対策に貢献する。