Syndromic testing panel for central nervous system infections using the LAMP method

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K07878
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52010:General internal medicine-related
• Research Institution: Fujita Health University
• Principal Investigator: Ihira Masaru 藤田医科大学, 保健学研究科, 教授 (10290165)
• Co-Investigator(Kenkyū-buntansha): 東本 祐紀 藤田医科大学, 医療科学部, 講師 (20569701) ; 吉川 哲史 藤田医科大学, 医学部, 教授 (80288472)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: Syndromic test / LAMP / real time PCR / HSV-1 / HSV-2 / VZV / HHV-6

Outline of Final Research Achievements

The LAMP method consists of a simple instrument structure, and the combination of microfluidic chips, which enables system miniaturization and multiplexing of tests, is suitable for the Syndromic testing panel. Comparison of LAMP and real-time PCR assays of spinal fluid suspected of encephalitis/encephalopathy showed concordance for HHV-6 and VZV LAMPs but did not have enough sensitivity to detect HSV-2 and EBV DNA by LAMPs. After determining the lower detection limit of multiplex LAMP-based on a microfluidic chip, the clinical utility was comfited using 32 sab samples. If real-time PCR was used as a standard, sensitivity of HSV-1, HSV-2, and VZV LAMPs based on microfluidic chip were 90%, 100%, and 100%, and the specificity of three methods were 96%, 100% and 100%, respectively. Multiplex LAMP, based on a microfluidic chip that detects three alpha herpesviruses, may be useful in laboratory diagnosis of patients with vesicular skin lesions.

Free Research Field

臨床ウイルス

Academic Significance and Societal Importance of the Research Achievements

感染症領域では、症状に基づき複数病原を同時に検査するsyndromic testingが行われPCRを基礎としている。本邦で開発されたLAMP法は、迅速性・簡便性・コストのアドバンテージがあり、LAMP法を基礎とするsyndromic testing構築の意義は大きい。本研究は、脳炎・脳症を対象とした場合、一部が不完全でありさらに検討を必要とするが、皮膚ぬぐい液による水疱性疾患識別のためのMicro流体LAMPについてはreal-timePCRと比較しても十分な感度を確認した。