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Mornitoring and Identification of Human Astrovirus from Groundwater in Korea Based on Highly Sensitive RT-nested PCR Primer Sets

  • Lee, Siwon (R&D Team, LSLK Co. Ltd.) ;
  • Bae, Kyung Seon (Water Supply and Sewerage Research Division, National Institute of Environmental Research) ;
  • Park, Jihyun (Water Supply and Sewerage Research Division, National Institute of Environmental Research) ;
  • Kim, Jin-Ho (Institute of Tissue Regeneration Engineering (ITREN), Dankook University) ;
  • Lee, Jin-Young (R&D Team, LSLK Co. Ltd.) ;
  • Choi, Jiwon (R&D Team, LSLK Co. Ltd.) ;
  • Park, Eung-Roh (Water Supply and Sewerage Research Division, National Institute of Environmental Research) ;
  • You, Kyung-A (Water Supply and Sewerage Research Division, National Institute of Environmental Research)
  • Received : 2021.09.16
  • Accepted : 2021.11.15
  • Published : 2021.12.31

Abstract

Human Astrovirus (HuAstV) is an important gastrointestinal pathogen that is frequently reported worldwide. Monitoring of contaminated groundwater has been suggested since HuAstV is transmitted through the fecal-oral route. This study developed a test method based on conventional reverse transcription (RT)-nested polymerase chain reaction (PCR) that involves SL® non-specific reaction inhibitor for unknown non-specific amplification taking place in the groundwater environment. An optimal method for detecting HuAstV in groundwater sample through analysis and comparison against conventionally reported method was also suggested. The developed method enabled the production of nested PCR amplicon of 630 nt, which is a sufficient length for similarity analysis based on sequencing and genotyping. Amplicons suspected to be HuAstV were amplified in two out of the twenty groundwater samples collected in Korea, presenting 99.77% and 99.73% similarity against HuAstV 1 strain lhar/2011/kor (JN887820.1) in sequencing, respectively. These amplicons were identified as HuAstV 1.

Keywords

Acknowledgement

This work was supported by a grant from the National Institute of Environmental Research (NIER), funded by the Ministry of Environment (MOE) of the Republic of Korea (NIER-2020-01-01-003).

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