{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T08:46:50Z","timestamp":1768985210567,"version":"3.49.0"},"reference-count":51,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T00:00:00Z","timestamp":1753833600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institute of Environmental Research (NIER)","award":["NIER-2023-01-01-166"],"award-info":[{"award-number":["NIER-2023-01-01-166"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hepatitis E virus (HEV) is a leading cause of acute viral hepatitis and is primarily transmitted via contaminated water and food. Groundwater may also serve as a potential vector for HEV transmission. This study aimed to optimize real-time polymerase chain reaction (rtPCR) for the detection of HEV, employing both TaqMan probe- and SYBR Green-based methods. A total of 12 primer sets for the TaqMan probe-based method and 41 primer sets for the SYBR Green-based method were evaluated in order to identify the optimal combinations. Primer sets #4 (TaqMan probe-based) and #21 (SYBR Green-based) demonstrated the highest sensitivity and specificity, successfully detecting HEV in artificially spiked samples at 1 fg\/\u03bcL. The TaqMan probe-based method facilitated rapid detection with minimized non-specific amplification, whereas the SYBR Green-based method allowed for broader primer exploration by leveraging melting curve analysis. Despite the absence of HEV detection in 123 groundwater samples, this study underscores the value of real-time PCR for environmental monitoring and paves the way for enhanced diagnostic tools for public health applications.<\/jats:p>","DOI":"10.3390\/ijms26157377","type":"journal-article","created":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T14:00:41Z","timestamp":1753884041000},"page":"7377","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Real-Time PCR-Based Detection of Hepatitis E Virus in Groundwater: Primer Performance and Method Validation"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8775-9102","authenticated-orcid":false,"given":"Jin-Ho","family":"Kim","sequence":"first","affiliation":[{"name":"Biomedical Research Institute, Dankook University Hospital, Cheonan 31116, Republic of Korea"},{"name":"Department of Chemistry, College of Science and Engineering, Dankook University, Cheonan 31116, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siwon","family":"Lee","sequence":"additional","affiliation":[{"name":"R&D Team, LSLK Co., Ltd., Gimpo 10111, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eung-Roh","family":"Park","sequence":"additional","affiliation":[{"name":"Water Use and Management Research Division, Water Environmental Research Department, National Institute of Environmental Research, Incheon 22689, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1907","DOI":"10.1016\/j.csbj.2021.03.038","article-title":"Structural and molecular biology of hepatitis E virus","volume":"19","author":"Wang","year":"2021","journal-title":"Comput. 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