{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T19:07:21Z","timestamp":1786734441677,"version":"build-2736575974"},"reference-count":58,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T00:00:00Z","timestamp":1779321600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100013885","name":"National Institutes of Health Malaysia","doi-asserted-by":"publisher","award":["NMRR-22-00823"],"award-info":[{"award-number":["NMRR-22-00823"]}],"id":[{"id":"10.13039\/501100013885","id-type":"DOI","asserted-by":"publisher"}]},{"award":["NMRR-22-00823"],"award-info":[{"award-number":["NMRR-22-00823"]}],"id":[{"id":"https:\/\/ror.org\/045p44t13","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Background: Wastewater-based surveillance (WBS) has emerged as a valuable tool for population-level monitoring of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) transmission, yet the interplay between sampling strategies and disease prevalence in shaping detection performance remains ambiguous. We investigated how grab and composite sampling influence SARS-CoV-2 ribonucleic acid (RNA) detection dynamics and predictive lag times across high- and low-prevalence communities in Selangor, Malaysia. Methods: A 28-week longitudinal study was conducted in Selangor, Malaysia, comparing grab and composite wastewater sampling in communities with high and low Coronavirus disease 2019 (COVID-19) prevalence. SARS-CoV-2 RNA in 348 samples was quantified using digital Reverse Transcription Polymerase Chain Reaction (RT-dPCR), and viral lineages were characterized by Nanopore sequencing. Detection sensitivity and lead times relative to reported cases were evaluated. Results: In low-prevalence settings, grab sampling showed higher detection sensitivity than composite sampling (92.0% vs. 70.0%), whereas both methods achieved similarly high detection in high-prevalence areas (&gt;97.0%). Lag-time analysis indicated that grab sampling in high-prevalence settings was significantly associated with case trends at potential two-week lead (p = 0.024), while composite sampling in low-prevalence settings showed the strongest association at a potential one-week lead (p = 0.0022). Overall, lag structures varied by both sampling strategy and prevalence context. Both sampling approaches captured the replacement of Omicron sublineages (XBB.1.5, XBB.1.9.1, XBB.1.16) and identified additional circulating variants, including EG.5, that were not captured in the available clinical sequencing dataset during the same period. Conclusions: These findings reveal that local transmission intensity is associated with the utility of different sampling designs. Context-specific optimization of WBS sampling strategies enhances sensitivity, reduces detection lag, and strengthens early warning and genomic-tracking capacity in public health surveillance frameworks.<\/jats:p>","DOI":"10.3390\/v18050583","type":"journal-article","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T15:02:27Z","timestamp":1779375747000},"page":"583","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Influence of Sampling Strategies and Disease Prevalence on SARS-CoV-2 Detection Dynamics in Wastewater Surveillance"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6609-1529","authenticated-orcid":false,"given":"Siti Aishah","family":"Rashid","sequence":"first","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohd Ishtiaq","family":"Anasir","sequence":"additional","affiliation":[{"name":"Infectious Disease Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fadly Syah","family":"Arsad","sequence":"additional","affiliation":[{"name":"Kuala Lumpur & Putrajaya Health Department, Lembah Pantai District Health Office, Kuala Lumpur 50480, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nurul Farehah","family":"Shahrir","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khayri Azizi","family":"Kamel","sequence":"additional","affiliation":[{"name":"Infectious Disease Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2260-8821","authenticated-orcid":false,"given":"Sakshaleni","family":"Rajendiran","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9704-5026","authenticated-orcid":false,"given":"Nurul Amalina","family":"Khairul Hasni","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9879-0642","authenticated-orcid":false,"given":"Mohamad Iqbal","family":"Mazeli","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4759-1840","authenticated-orcid":false,"given":"Yuvaneswary","family":"Veloo","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8743-7154","authenticated-orcid":false,"given":"Syahidiah","family":"Syed Abu Thahir","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan Rozita","family":"Wan Mahiyuddin","sequence":"additional","affiliation":[{"name":"SEAMEO TROPMED Network Malaysia, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6739-652X","authenticated-orcid":false,"given":"Khor Bee","family":"Chin","sequence":"additional","affiliation":[{"name":"Indah Water Konsortium Sdn. Bhd., Kuala Lumpur 60000, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alijah","family":"Mohd Aris","sequence":"additional","affiliation":[{"name":"Indah Water Konsortium Sdn. Bhd., Kuala Lumpur 60000, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Redzuan","family":"Zainudin","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8811-5843","authenticated-orcid":false,"given":"Rafiza","family":"Shaharudin","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3219-4204","authenticated-orcid":false,"given":"Raheel","family":"Nazakat","sequence":"additional","affiliation":[{"name":"Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam 40170, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,21]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2026, January 13). 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