Lab Matters Fall 2022 | Page 39

APHL 2022 POSTER ABSTRACTS nucleic acid is then processed using either a spin column or using an automated nucleic acid purification system . The process recovered 63.13 ± 4.16 %, 40.09 ± 10.89 %, 39.67 ± 10.66 %, of HCoV OC43 , 229E and bacteriophage MS2 . Using this workflow , we monitored wastewater samples from three WWTP in Dane County , Wisconsin , serving a combined population of over 300,000 people , for one year . As wastewater often contains RT-qPCR inhibitors , we formulated enzyme mixes that demonstrated resistance to PCR inhibitors commonly found in wastewater such as humic acid and urea . We developed a SARS-CoV-2 RT-qPCR assay using US CDC N1 , N2 ( nucleocapsid ) and the E ( envelope ) gene fragments . Pepper Mild Mottle Virus ( PMMoV ), a fecal indicator RNA virus present in wastewater and used as a normalization tool . SARS-CoV-2 variants of concerns ( alpha , beta , gamma , delta ) and enteric viruses ( Norovirus GI , GII and Hepatitis A ) were also analyzed using the method . The workflow has a limit of detection of 1.6 GC / ml with 40 ml of raw wastewater sample for SARS-CoV-2 . Our analysis was compared to the COVID-19 cases declared by the municipalities , showed strong correlation between the SARS-CoV-2 viral load present in wastewater and clinical cases . The alpha and delta variant started appearing in wastewater samples in March and May , 2021 respectively . Norovirus GII was detected in all samples and GI in 25 % of the samples tested . HepA was not detected in any sample .
Presenter : Subhanjan Mondal , Promega Corporation , subhanjan . mondal @ promega . com
Intelligent Routing and Challenges of COVID-19 Electronic Lab Reporting and Applicability to Future Pandemics
M Malai 1 , C Simpson 2 , B Garrett 1 , P Zarcone-Gagne 1 , M Gibbons 1 ;
1
Association of Public Health Laboratory , 2 AIMS + / APHL / Ruvos
As COVID-19 breaking news began to flood our media , we were thrown into the midst of a pandemic and teams across the country pulled together to strategize and take action . The US Department of Health and Human Services ( HHS ) released guidance to ensure appropriate COVID-19 Electronic Lab Reporting ( ELR ) data reached Public Health Agencies ( PHAs ), requiring all testing facilities to comply with the requirements found in the CARES Act . One issue in particular that was becoming increasingly transparent was the lack of accessible infrastructure , data transport and data quality validation for private laboratories that found themselves in the COVID-19 testing space . In response to the challenges surrounding COVID-19 test results reporting , the Association of Public Health Laboratories ( APHL ) launched the COVID-19 Centralized ELR project utilizing the AIMS Platform ( AIMS ). AIMS is a secure , FISMAcompliant , cloud-based environment that is widely-adopted by state , local and federal public health stakeholders . These factors accelerated getting higher-quality COVID-19 ELR data exactly where it needed to be received . For mandated ELR to PHAs such as COVID-19 , AIMS acts as a hub , allowing the sender to transmit ELR messages to a single connection where AIMS securely receives , routes and delivers this data to PHAs through already-established connections . This single connection significantly decreased the time-to-production from months to weeks and alleviates burden by reducing the reporting burden on testing laboratories , leveraging the existing AIMS infrastructure and connections , facilitating collaboration and communication with PHAs , facilitating quality control and COVID-19 standards preservation , and reducing point-topoint connections with PHAs . APHL ’ s Centralized ELR project allows private laboratories to deliver secure , critical SARS-CoV-2 results to 55 PHAs . The project is in a continuous state of onboarding new COVID-19 Centralized ELR reporters , facilitating communication and improvements among reporters and PHAs , and addressing challenges that have come-up along the way like multiplex , variant and school-based COVID-19 test results reporting . We will share information on the Centralized ELR intelligent data flow , challenges encountered , solutions to address those challenges , discuss impact and benefits to public health , and metrics quantifying the scope and scale of that impact .
Presenter : Caitlyn Simpson , Ruvos / AIMS +/ APHL , Email : csimpson @ ruvos . com
Demonstration of the Usefulness of a Quantitative SARS CoV-2 Human IgG Multiplex Antibody Assay
K Venkateswaran , N Venkateswaran , J Walker , J Sarwar and W Nelson , Tetracore , Inc .
Understanding the kinetics of antibody response to SARS-CoV-2 ( CoV2 ) after infection and vaccination is critical to assess the utility of serological testing at individual and population levels during the COVID 19 pandemic , which continues to cause a significant burden on global public health . Several serological tests available today provide a qualitative result for the presence or absence of antibody reactivity to a single antigen in a sample . Although CoV2 is a novel coronavirus causing severe respiratory illness , four common cold human coronaviruses ( HCoV 229E , OC43 , NL-63 and HKU1 ) have circulated in the human population for a prolonged duration and have become endemic . We developed multiplex assays based on Luminex ® xMAP ® technology to estimate circulating IgG antibodies to multiple recombinant proteins from wild-type and variant CoV-2 , SARS-CoV , MERS-CoV and four other HCoVs . We evaluated over 200 samples following infection , 50 samples after vaccination and 168 samples from PCR negative or normal healthy individuals . A qualitative 7-plex assay used three wild-type CoV-2 specific antigens — Receptor Binding Domain ( RBD ), Nucleocapsid Protein ( NP ) and a hybrid RBD-NP protein . Quantitative human IgG antibody response to three CoV2 recombinant antigens , RBD , NP and trimeric spike protein , was tested in these samples using WHO quantification standard . Higher levels of antibodies were found in vaccinated samples compared to the infection . We performed analytical validation of the multiplex quantitative IgG antibody assay to determine its performance characteristics . Results showed that multiplex quantitative antibody assay could be used very effectively for measuring immune responses to SARS-CoV-2 antigens after infection or vaccination . The multiplex assay has great potential to differentiate immune response induced by natural infection from vaccination . Multiplex quantitative antibody assay can serve as a surrogate test to indicate correlates of protection following SARS- CoV-2 infection and vaccination .
Presenter : Neeraja Venkateswaran , Tetracore , Inc ., nvenkateswaran @ tetracore . com
COVID-19
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