2021 Fiscal Year Final Research Report

Research on establishment of rapid microbial identification method for various forensic samples using nanopore sequencer

Research Project

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Project/Area Number	19K10703
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Review Section	Basic Section 58040:Forensics medicine-related
Research Institution	National Research Institute of Police Science
Principal Investigator	Muto Junji 科学警察研究所, 法科学第一部, 主任研究官 (80432186)
Co-Investigator(Kenkyū-buntansha)	堀田明豊国立感染症研究所, 獣医科学部, 主任研究官 (90392323)
Project Period (FY)	2019-04-01 – 2022-03-31
Keywords	ナノポアシーケンサー / 微生物 / ロングリード / 検出 / 感染臓器 / ゲノム解析 / 同定 / 法科学的試料
Outline of Final Research Achievements	We investigated whether the reads of Francisella tularensis could be detected from the organs and blood clots of the hares dead from tularemia using a nanopore sequencer. It was difficult to isolate and culture F. tularensis from the samples of the rotting carcass after death. However, the reads of F. tularensis could be detected from these samples using a nanopore sequencer. On the other hand, a detailed analysis of the reads of the samples revealed that the read length was short. Therefore, we changed the sample preparation to the method suitable for obtaining long reads. Preparation of the blood clots of the rotting carcass by this method increased read length. In addition, the Streptococcus pyogenes-contained blood sample, which had been frozen and stored for 11 years, was prepared by this method, and it was examined whether the reads of S. pyogenes could be detected in the same manner. As a result, the reads of S. pyogenes could be detected.
Free Research Field	微生物学、ウイルス学、法医学
Academic Significance and Societal Importance of the Research Achievements	食品汚染や生物テロ等の病原微生物を用いた犯罪では、被害者の処置、被害拡大防止の対応および捜査のため、試料に含まれる微生物を迅速に検出・同定することが求められる。本研究では、迅速性や利便性で注目されているナノポアシーケンサーを用いて、血液や臓器等の法科学的試料から含まれている微生物を検出できるか検証した。野兎病で死亡したノウサギ死体の臓器や化膿レンサ球菌を含む血液試料から目的の細菌を検出でき、法科学的試料から微生物を検出・同定するのにナノポアシーケンサーは有用と考えられる。