{"id":14082,"date":"2022-04-08T16:50:41","date_gmt":"2022-04-08T20:50:41","guid":{"rendered":"https:\/\/www.vdh.virginia.gov\/environmental-health\/?page_id=14082"},"modified":"2022-04-08T17:12:05","modified_gmt":"2022-04-08T21:12:05","slug":"sample-collection-and-laboratory-methods","status":"publish","type":"page","link":"https:\/\/www.vdh.virginia.gov\/environmental-health\/wastewater-surveillance-for-covid-19\/sample-collection-and-laboratory-methods\/","title":{"rendered":"Sample Collection and Laboratory Methods"},"content":{"rendered":"<p><span style=\"font-weight: 400\">Sample collection is done by the participating utilities.\u00a0 This program is reliant upon our utility partners for the proper collection, handling, and storage of the sample until the sample is picked up for transport to the laboratory, DCLS.\u00a0\u00a0<\/span><\/p>\n<p><span style=\"font-weight: 400\">There are two main types of samples that are collected:\u00a0 grab samples and composite samples.\u00a0\u00a0<\/span><\/p>\n<ul>\n<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Grab samples are a single sample collected at a moment in time. This type of sample is representative of the wastewater flowing into the treatment plant at that moment the sample is collected.<\/span><\/li>\n<li style=\"font-weight: 400\"><span style=\"font-weight: 400\">Composite samples are individual samples collected over a 24-hour time period and then combined to form a single sample.\u00a0 The samples can be collected based on a set time (for example, every 10 minutes) or depending on how much flow enters the wastewater treatment plant.\u00a0 This type of sample is representative of the whole day of flow into the wastewater treatment plant.<\/span><\/li>\n<\/ul>\n<p><span style=\"font-weight: 400\">The utility provides additional information (metadata) that accompanies the sample such as time of sample collection, the flow for the day of collection, and the sampling point.\u00a0 The samples are all collected at the entrance to the treatment plant before any significant processing occurs.<\/span><\/p>\n<p><span style=\"font-weight: 400\">Samples are shipped to DCLS for concentration, extraction, and droplet digital polymerase chain reaction (ddPCR) analysis. Prior to the week of November 15, 2021, extracted samples were shipped to the NELAP-Accredited laboratory at Hampton Roads Sanitation District (HRSD) for SARS-CoV-2 analysis via ddPCR. <\/span><span style=\"font-weight: 400\">The gene target presented in this report is ddCoV_N, a modified version of the N2 gene for ddPCR wastewater analysis<\/span><span style=\"font-weight: 400\">[1]<\/span><span style=\"font-weight: 400\">. <\/span><span style=\"font-weight: 400\">Quantification results for which amplification was observed in the no-template controls (NTC) are removed from analysis. The limit of detection for the current version of the laboratory protocol (v2.0) is 6,200 copies\/L of wastewater. All concentrations below the limit of detection are assigned a value equal to one half the limit of detection.\u00a0<\/span><\/p>\n<p>&nbsp;<\/p>\n<table style=\"height: 212px;border-color: #000000\" width=\"943\">\n<tbody>\n<tr style=\"border-color: #000000\">\n<td><b>Lab Protocol Version<\/b><\/td>\n<td><b>Limit of Detection (LOD)<\/b><\/td>\n<td><b>Laboratory Responsibilities<\/b><\/td>\n<\/tr>\n<tr>\n<td style=\"border-color: #000000\"><span style=\"font-weight: 400\">v2.0<\/span><\/td>\n<td><span style=\"font-weight: 400\">6,200<\/span><\/td>\n<td><b>Concentration, extraction, and ddPCR analysis: <\/b><span style=\"font-weight: 400\">DCLS<\/span><\/td>\n<\/tr>\n<tr>\n<td style=\"border-color: #000000\"><span style=\"font-weight: 400\">v1.0<\/span><\/td>\n<td><span style=\"font-weight: 400\">1,350<\/span><\/td>\n<td><b>Concentration and extraction: <\/b><span style=\"font-weight: 400\">DCLS<\/span><\/p>\n<p><b>Droplet digital polymerase chain reaction (ddPCR) analysis: <\/b><span style=\"font-weight: 400\">HRSD<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<p><span style=\"font-weight: 400\">Viral load is calculated based on Equation 1:<\/span><\/p>\n<p><span style=\"font-weight: 400\">(1) Viral load (copies\/day) = Viral conc. (copies\/L)\u2217Flow rate (MGD)\u2217(3785411.8 L\/d)\/(1 MGD)<\/span><\/p>\n<p>&nbsp;<\/p>\n<p><span style=\"font-weight: 400\">Percentiles are calculated on a per-site basis by viral load, and then binned into quintiles (i.e., 0-19.9th, 20-39.9th, 40-59.9th, 60-79.9th, 80-100th). Site-specific percentiles provide a metric to compare the current week&#8217;s data to the site&#8217;s historical data, which allows for color visualization of when each site experienced its highest versus lowest loads.<\/span><\/p>\n<p><b>References<\/b><\/p>\n<p><span style=\"font-weight: 400\">[1] Gonzalez, R., Larson, A., Thompson, H., Carter, E., &amp; Cassi, X. F. (2021). Redesigning SARS-CoV-2 clinical RT-qPCR assays for wastewater RT-ddPCR. https:\/\/doi.org\/10.1101\/2021.03.02.21252754<\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Sample collection is done by the participating utilities.\u00a0 This program is reliant upon our utility partners for the proper collection, handling, and storage of the sample until the sample is picked up for transport to the laboratory, DCLS.\u00a0\u00a0 There are two main types of samples that are collected:\u00a0 grab samples and composite samples.\u00a0\u00a0 Grab samples [&hellip;]<\/p>\n","protected":false},"author":404,"featured_media":0,"parent":10885,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_lmt_disableupdate":"","_lmt_disable":"","footnotes":""},"tags":[],"class_list":["post-14082","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.4 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Sample Collection and Laboratory Methods - 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