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2021 年度 実施状況報告書

Molecular characterization, its mutation pattern and the molecular surveillance of SARS-CoV-2 in Hiroshima

研究課題

研究課題/領域番号 21K21113
研究機関広島大学

研究代表者

KO KO  広島大学, 医系科学研究科(医), 助教 (80910299)

研究期間 (年度) 2021-08-30 – 2023-03-31
キーワードSARS-CoV-2 / Variants screening / Sanger method
研究実績の概要

Total 239 saliva samples from the confirmed cases of COVID-19 collected from 2020.12.1 to 2022.3.14 in Hiroshima were screened for SARS-CoV-2 variants using Sanger based sequencing method. 159 out of 239 samples had been amplified using in-house primer set targeting the S1 spike gene of SARS-CoV-2 (22,735nt~23,532nt) and the variants were identified through the unique checkpoints in the targeted gene. The viral titer ranges from 8 to 5E+8 copies/mL with an average of 8E+6 copies/mL. There is no significant difference in viral titer by variants. No mutation at spike gene was 16 (10%), E484K mutation (R.1) was 3 (1.9%), Alpha (B.1.1.7) was 33 (20.8%), Delta (B.1.627) was 65 (40.9%) and Omicron (B.1.1.529) was 42 (26.4%). The remaining 80 samples were now ongoing for the variant screening.

現在までの達成度 (区分)
現在までの達成度 (区分)

2: おおむね順調に進展している

理由

159 out of 239 samples from confirmed cases of COVID-19 in Hiroshima had been performed Sanger based variant screening targeting the S1 spike gene of SARS-CoV-2. 66.5% of samples were completed for variant screening. Now, the amplification of remaining 80 samples were completed and now proceed to sequencing. 40% of remaining work had been completed.

今後の研究の推進方策

The remaining 80 samples were ongoing for the variants screening. Then, the genomes data will be analyzed using Genetynx Mac and the validation of Sanger sequencing will be performed by homology analysis of genomes data from Sanger and Next generation Sequencing of each single sample. After finalizing the result, the complete report will be done.

次年度使用額が生じた理由

The variant screening of samples included this study was performed using the existing reagents and primers of another project. Therefore, the usage of this grant is less than expected. But, from now, the new reagents and other laboratory equipment will be bought in order to continue the research.

  • 研究成果

    (1件)

すべて 2022

すべて 雑誌論文 (1件) (うち国際共著 1件、 査読あり 1件、 オープンアクセス 1件)

  • [雑誌論文] Mass Screening of SARS-CoV-2 Variants using Sanger Sequencing Strategy in Hiroshima, Japan2022

    • 著者名/発表者名
      K Ko, K Takahashi, S Nagashima, B E, S Ouoba, MRA Hussain, T Akita, A Sugiyama, T Sakaguchi, H Tahara, H Ohge, H Ohdan, T Kubo, N Ishikawa, T Takafuta, Y Fujii, M Mimori, F Okada, E Kishita, K Ariyoshi, M Kuwabara, J Tanaka
    • 雑誌名

      Scientific Reports

      巻: 12 ページ: 2419

    • DOI

      10.1038/s41598-022-04952-2

    • 査読あり / オープンアクセス / 国際共著

URL: 

公開日: 2022-12-28  

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