Lab Matters Fall 2023 | Page 60

APHL 2023 POSTER ABSTRACTS

detection from human and animal nasopharyngeal swab samples The SERS sensor will be prepared based on a silver nanorod array ( AgNR ) substrate by assembling probe DNA to capture SARS-CoV-2 RNA . SERS spectra of nasopharyngeal specimens from positive and negative will be identified in less than 30 minutes . The probe DNA functionalized AgNR array SERS sensor combining with deep learning could serve as a potential rapid point-of-care COVID-19 diagnostic platform for testing of humans and animal samples .

Presenter : Teddy Spikes , trs53790 @ uga . edu

Reduced Ion Suppression in an Automated Extraction of Vitamins B1 and B6 from Whole Blood for LCMS Analysis

K . Dukes ; Biotage

Phospholipids and proteins in blood and plasma sample are a main cause of ion suppression in liquid chromatography-mass spectrometry ( LCMS ) analysis . In addition to ion suppression , the phospholipids and proteins in blood and plasma samples can cause premature column failures and increase frequency of instrument maintenance . Presented is the work demonstrating the effectiveness of the Biotage ISOLUTE PLD + ™ phospholipid removal plate used in the extraction of Vitamins B1 and B6 from human whole blood . The PLD + plate can be incorporated into an existing method with little or no modification , as the blood or plasma is precipitated directly on the plate and can be easily automated . The Biotage ISOLUTE PLD + ™ demonstrated an over 90 % reduction in phospholipids and proteins and an increase in signal-to-noise ratio of the endogenous levels of B1 and B6 .

Presenter : Kyle Dukes , kyle . dukes @ biotage . com

The Development of a Multiplexed Immunoassay for Multi- Stage Lyme Disease Detection

J . Needham , D . Reyes , G . Ireton , A . Vallur ; InBios International , Inc .

In the United States , Lyme disease is primarily caused by Borrelia burgdorferi sensu stricto , spread via the black-legged tick , Ixodes scapularis . Symptoms of early Lyme disease may include fever , rash , headache and fatigue . Later stages of Lyme disease , particularly if left untreated , may lead to complications including arthritis , memory loss and carditis . Lyme disease remains the most widely spread vector-borne disease in North America with approximately 476,000 infections in the US each year causing an annual economic burden approaching $ 1 billion ; however , the accurate and early diagnosis for Lyme disease infection remains challenging . The low parasite load and early dissemination of the spirochete limit the sensitivities for direct detection assays . Discernable manifestations , such as erythema migrans ( EM ), may only present in up to ~ 70 % of patients . At this time , serological assays remain the mainstay of laboratory confirmation though it may take weeks to develop a sufficient antibody response . The US Centers for Disease Control ( CDC ) recommends testing sera with a “ standard ” or “ modified ” two-tier testing algorithm ( STTA or MTTA ) which requires testing a specimen with a sensitive immunoassay followed by either an immunoblot or enzyme-linked immunoassay ( ELISA ) to confirm the diagnosis . The development of a singular , more robust serological assay that can detect Lyme infection at both early and late stages of the disease will significantly improve clinical management and patient well-being . Here , we describe the development and preliminary evaluation of the InBios Lyme Detect™ Multiplex ELISA which uses a microarrayed panel of target proteins to simultaneously detect IgG and IgM reactivity independently to each antigen and interprets results through a machine learning ( ML ) algorithm to properly categorize a given specimen . This assay is performed using an ELISA format to ensure easy adaptation by clinical laboratories . Proprietary software is used to automatically analyze the captured images , determine signals for each target antigen and provide the ML interpretation for each tested specimen . Preliminary testing with 35 confirmed and well-characterized early Lyme ( EM positive ) disease specimens , including Panels 1 and 2 obtained from the CDC and the Lyme disease biobank of the Bay Area Lyme Society shows that the sensitivity for detecting early-stage infection is improved by approximately 37 % when compared to the STTA ( 48.6 % for STTA vs . 85.7 % for the Lyme DetectTM Multiplex ELISA ) while the specificity with other disease , endemic normal and nonendemic normal human serum samples is ≥98.5 %. Furthermore , independent reactivity for IgG and IgM is recorded for each target antigen , providing critically relevant data to the physician regarding the patient ’ s specific antibody response .

Presenter : James Needham , james @ inbios . com

The Role of Circulating Influenza Viruses on B Cell Memory

V . Vyas , Y . Wang , M . Sheth , M . Reed , J . Pohl , S . Gangappa , S . Sambhara , U . Shanmugasundaram , R . Kumari , M . Mishina , S . Sharma ; US Centers for Disease Control and Prevention

Influenza viruses cause annual seasonal epidemics and have the potential to cause occasional global pandemics due to antigenic shift . In the United States , influenza primarily circulates from August through April , except for the 2009 pandemic , which caused infections in early summer . Subclinical infections may stimulate the immune system and contribute to the persistence of memory post-vaccination . Before the COVID-19 pandemic , there was no way to address whether circulating influenza viruses contributed to the persistence of B cell memory . However , the COVID-19 pandemic presented a unique situation in which influenza circulation was not detectable during the 2020-2021 flu season . In addition , social distancing and wearing masks to mitigate the spread of SARS-CoV-2 also diminished the incidence of influenza , which allowed us to test if circulating influenza viruses impact influenza B cell memory . To address this question , we are developing a comprehensive immune monitoring workflow to characterize the frequencies of H3 hemagglutinin ( HA ) -specific B cells via high dimensional multiparametric flow cytometry , H3 HA-specific B cell receptor repertoire with 10X Genomics platform , serum H3 HA-specific IgG and IgA antibody repertoire by Liquid Chromatography and Mass Spectroscopy ( LC / MS ). After establishing the workflow , we will test the clinical samples to assess the impact of circulating viruses on B cell memory in individuals vaccinated in during the 2019-2020 and 2020-2021 influenza seasons .

Presenter : Ved Vyas , uao4 @ cdc . gov