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Development and Assessment of Specific and High Sensitivity Reverse Transcription Nested Polymerase Chain Reaction Method for the Detection of Aichivirus A Monitoring in Groundwater

지하수 중 Aichivirus A 모니터링을 위한 특이적 및 고감도 이중 역전사 중합효소연쇄반응 검출법 개발 및 평가

  • Bae, Kyung Seon (Water Supply and Sewerage Research Division, National Institute of Environmental Research) ;
  • Kim, Jin-Ho (Institute of Tissue Regeneration Engineering, Dankook University) ;
  • Lee, Siwon (R&D Team, LSLK Co.) ;
  • Lee, Jin-Young (R&D Team, LSLK Co.) ;
  • You, Kyung-A (Water Supply and Sewerage Research Division, National Institute of Environmental Research)
  • 배경선 (국립환경과학원 상하수도연구과) ;
  • 김진호 (단국대학교 조직재생공학연구원) ;
  • 이시원 (랩슬리코리아 R&D팀) ;
  • 이진영 (랩슬리코리아 R&D팀) ;
  • 유경아 (국립환경과학원 상하수도연구과)
  • Received : 2021.08.26
  • Accepted : 2021.09.16
  • Published : 2021.09.30

Abstract

Human Aichivirus (Aichivirus A; AiV-A) is a positive-sense single-strand RNA non-enveloped virus that has been detected worldwide in various water environments including sewage, river, surface, and ground over the past decade. To develop a method with excellent sensitivity and specificity for AiV-A diagnosis from water environments such as groundwater, a combination capable of reverse transcription (RT)-nested polymerase chain reaction (PCR) was developed based on existing reported and newly designed primers. A selective method was applied to evaluate domestic drinking groundwater samples. Thus, a procedure was devised to select and subsequently identify RT-nested PCR primer sets that can successfully detect and identify AiV-A from groundwater samples. The findings will contribute to developing a better monitoring system to detect AiV-A contamination in water environments such as groundwater.

사람 아이치바이러스(Aichivirus A; AiV-A)는 positivesense single-strand RNA 비외피 바이러스로 지난 10년 동안 하수, 강, 지표 및 지상의 다양한 물환경에서 전 세계적으로 검출이 보고되고 있다. 지하수 등 물환경에서 AiV-A 진단을 위한 고감도 및 특이성이 우수한 방법의 개발이 요구됨에 따라, 본 연구에서는 기존 및 신규 설계된 프라이머 세트를 기초로 역전사(RT) 및 이중 중합효소연쇄반응이 가능한 조합을 개발하였다. 개발한 방법을 국내 음용 지하수 시료에 적용 및 평가하였으며, 그 결과 지하수 시료에서 AiV-A를 성공적으로 검출 및 동정할 수 있는 RT-nested PCR primer sets가 선정되었고 후속적으로 동정할 수 있는 절차가 고안되었다. 본 연구 결과는 지하수 등 물 환경에서 AiV-A 오염을 탐지하기 위한 모니터링 시스템 마련에 기여할 것으로 기대된다.

Keywords

Acknowledgement

본 연구는 환경부의 재원으로 국립환경과학원의 지원을 받아 수행하였습니다(NIER-2020-01-01-003).

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