2021 Fiscal Year Final Research Report
Development of a highly sensitive detection method for Central Nervous System infections using Droplet Digital PCR
| Project/Area Number |
20K16534
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 52010:General internal medicine-related
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| Research Institution | Fujita Health University |
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Principal Investigator |
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| Project Period (FY) |
2020-04-01 – 2022-03-31
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| Keywords | ヘルペスウイルス / ddPCR |
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| Outline of Final Research Achievements |
Acute encephalitis/encephalopathy is a central nervous system infection with a poor prognosis that occurs in all age groups, but many issues remain, including the elucidation of the aetiology and pathogenesis. In the differential diagnosis, the cause is still unknown in about half of cases using the real-time PCR (qPCR) method, which is the current golden standard method, and a more sensitive method is desired. In this study, a comparison with existing qPCR methods was conducted to establish a method for detecting three types of α-herpesviruses using ddPCR methods. Three ddPCR methods did not provide better results than the current qPCR methods. There is room for improvement, such as increasing the amount of sample added to the ddPCR method or examining the appropriate annealing temperature for each virus detection system, in order to increase sensitivity.
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| Free Research Field |
感染症
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では、HSV-1とVZVはqPCR法がddPCR法より感度が良く、HSV-2ではddPCR法とqPCR法は同程度の感度であった。既報ではddPCR法がqPCR法よりも高感度との報告も多数あるため、さらなる改善の余地がある。既存より高感度分子生物学的診断法を構築することで、原因不明脳炎・脳症の中から新たに起因ウイルス同定が可能になり、それらがどのような臨床像を呈しているか明らかになる。また未診断例の中から新たにHSV、VZV陽性検体が見つかれば、早期診断、適切な抗ウイルス剤投与により患者予後の改善に直結する。
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