Development of rapid identification technique for pathogenic bacteria causing sepsis

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K09402
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 55060:Emergency medicine-related
• Research Institution: Nara Medical University
• Principal Investigator: Kawai Yasuyuki 奈良県立医科大学, 医学部, 助教 (90445073)
• Co-Investigator(Kenkyū-buntansha): 中山 章文 岐阜医療科学大学, 保健科学部, 教授 (70536721)
• Project Period (FY): 2019-04-01 – 2023-03-31
• Keywords: 敗血症 / 16SrRNA / 迅速診断

Outline of Final Research Achievements

We have designed unique primers targeting 23 bacterial species, which are the major causes of sepsis, while eliminating non-specific reactions with human DNA. Initially, we encountered a problem in detecting gram-positive bacteria, but after numerous revisions to the design and in silico analysis, we confirmed that each bacterial species could be amplified. We also verified that non-specific amplification reactions with human DNA could be suppressed.

Now, using these primers, we are analyzing samples taken from septicemic blood that tested positive in blood culture tests. We have discovered that both gram-negative and gram-positive bacteria, which pose significant clinical challenges, can be amplified and detected.

Free Research Field

救急集中治療

Academic Significance and Societal Importance of the Research Achievements

本研究では、敗血症の血液検体から原因菌を選択的に増幅するために、広い範囲の細菌を増幅が可能かつ、サンプル中に大量に含まれるヒトDNAとの非特異的増幅の排除するプライマーデザインとPCR条件の確立を行った。これらの成果によって敗血症血液検体から、これまで検査時間短縮のボトルネックとなる血液培養による増菌の過程を経ずに、直接細菌DNAを増幅可能となった。