Antigen-based Assay Research Articles

Correction: Sebro et al. Implementation of Antigen-Based Diagnostic Assays for Detection of Histoplasmosis and Cryptococcosis among Patients with Advanced HIV in Trinidad and Tobago: A Cross-Sectional Study. J. Fungi 2024, 10, 695

In the original publication [...]

Hapten Synthesis, Antibody Generation, and Immunoassay Development for Linezolid Therapeutic Monitoring.

Linezolid (LNZ) is a broad-spectrum antibiotic used for the treatment of severe Gram+ infections. Significant pharmacokinetic variability in different groups of patients requires therapeutic drug monitoring (TDM) to ensure optimal therapy. This article presents the first description of the development of immunoanalytical TDM systems for LNZ. Four LNZ derivatives (H1-H4) with spacer arms of a different length (0-6.2-9.4-11.8 Å) and chemistry (aliphatic and heterocyclic) were synthesized to prepare H1-H3-immunogens and H1/H4-coating antigens or H1/H2-enzyme tracers. Three distinct antibody types against H1-H3 haptens were generated in rabbits and served as the basis for the development of ELISAs in heterologous coated antigen and coated antibody formats. The length of the spacer in the immunogen had an insignificant effect on the quality of the antibody generated or the performance of the designed coated antibody-ELISA formats (IC50 = 0.48-0.75 ng/mL) and coated antigen ELISAs (IC50 = 11.5-28.3 ng/mL). The coated antigen-based assays were considered more appropriate for monitoring therapeutic levels of LNZ due to their more convenient reduced sensitivity but wider dynamic range than the coated antibody-based assays (2.0-160 vs 0.16-2.1 ng/mL). The best recovery of LNZ (82.1-99.6%) from spiked human serum among the sample pretreatments examined was provided by the trichloroacetic acid deproteinization procedure. Serum samples derived from surgical patients with superimposed Gram-positive infections treated with LNZ were evaluated by parallel ELISAs using different immunoreagents. The results obtained in the various assays were found to be concordant, thereby confirming the reliability of the measurements and the developed assay systems suitable for TDM purposes.

Implementation of Antigen-Based Diagnostic Assays for Detection of Histoplasmosis and Cryptococcosis among Patients with Advanced HIV in Trinidad and Tobago: A Cross-Sectional Study.

The Caribbean continues to have high HIV prevalence globally with concurrently high mortality rates due to opportunistic Infections. This study addresses the prevalence of histoplasmosis and cryptococcosis among patients living with advanced HIV disease (AHD) in Trinidad and Tobago, focusing on the implementation of antigen-based diagnostic assays. Conducted as a cross-sectional survey across five HIV treatment sites, 199 participants with advanced HIV disease were enrolled between July 2022 and September 2023. Diagnostic testing was performed using the Clarus Histoplasma Galactomannan Enzyme Immunoassay (EIA), and the Immy CrAg® LFA Cryptococcal Antigen Lateral Flow Assay on urine and blood samples, respectively. Results revealed that 14.6% of participants were found to be co-infected with either histoplasmosis or cryptococcosis, with histoplasmosis being more prevalent (10.5%) than cryptococcosis (4.0%). The study found no significant demographic differences between newly diagnosed and previously diagnosed participants. However, a lower median CD4 count was associated with a higher risk of fungal opportunistic infections. The findings underscore the critical role of systematic use of fungal antigen-based diagnostic assays among patients with AHD to improve the timely diagnosis and treatment of fungal infections among people living with HIV in resource-limited settings and to improve patient outcomes and survival.

Ultrafast and highly sensitive detection of SARS-CoV-2 spike protein by field-effect transistor graphene-based biosensors

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), etiological agent for the coronavirus disease 2019 (COVID-19), has resulted in over 775 million global infections. Early diagnosis remains pivotal for effective epidemiological surveillance despite the availability of vaccines. Antigen-based assays are advantageous for early COVID-19 detection due to their simplicity, cost-effectiveness, and suitability for point-of-care testing (PoCT). This study introduces a graphene field-effect transistor-based biosensor designed for high sensitivity and rapid response to the SARS-CoV-2 spike protein. By functionalizing graphene with monoclonal antibodies and applying short-duration gate voltage pulses, we achieve selective detection of the viral spike protein in human serum within 100 µs and at concentrations as low as 1 fg ml−1, equivalent to 8 antigen molecules per µl of blood. Furthermore, the biosensor estimates spike protein concentrations in serum from COVID-19 patients. Our platform demonstrates potential for next-generation PoCT antigen assays, promising fast and sensitive diagnostics for COVID-19 and other infectious diseases.

Evaluation of Anti-CAR Linker mAbs for CAR T Monitoring after BiTEs/bsAbs and CAR T-Cell Pretreatment.

For the monitoring of chimeric antigen receptor (CAR) T-cell therapies, antigen-based CAR detection methods are usually applied. However, for each target-antigen, a separate detection system is required. Furthermore, when monitored CAR T-cells in the blood of patients treated with bispecific antibodies or T-cell engagers (bsAbs/BiTEs) recognize the same antigen, these methods produce false-positive results in clinical diagnostics. Anti-CAR-linker monoclonal antibodies (mAbs) targeting the linker sequence between the variable domains of the antigen binding CAR fragment promise a universal and unbiased CAR detection. To test this, we analyzed clinical specimens of all BCMA- and CD19-targeting CAR T-cell products currently approved for clinical use. We found a highly specific and sensitive CAR detection using anti-CAR-linker mAb in blood cells from patients treated with Ide-cel, Tisa-cel, Axi-cel, Brexu-cel, and Liso-cel. For Ide-cel and Tisa-cel, the sensitivity was significantly lower compared to that for antigen-based CAR detection assays. Strikingly, the specificity of anti-CAR linker mAb was not affected by the simultaneous presence of bispecific blinatumomab or teclistamab for Axi-cel, Brexu-cel, Liso-cel, or Ide-cel, respectively. Cilta-cel (containing a monomeric G4S-CAR linker) could not be detected by anti-CAR linker mAb. In conclusion, anti-CAR-linker mAbs are highly specific and useful for CAR T-cell monitoring but are not universally applicable.

Recombinant 3AB3 nonstructural protein-based indirect ELISA for detection of foot-and-mouth disease virus infection-elicited antibodies in goat.

Foot-and-mouth disease (FMD) is one of the most important animal diseases of economic significance globally. It is a highly infectious and contagious disease of cloven-hoofed animals including sheep and goat. For sero-diagnosis of FMD, recombinant antigen-based assays are considered as alternatives to conventional approaches such as the liquid phase blocking ELISA (LPBE). The early interventions towards control measures cannot be implemented unless the disease gets promptly diagnosed. It is relatively difficult to clinically diagnose FMD in goat due to the usual milder form or unapparent nature of symptoms. Under such situations where clinical samples are not available, demonstration of infection-specific FMD virus (FMDV) antibodies in serum sample may help identifying the animals exposed to the virus in retrospect. Antibody to 3AB nonstructural protein (NSP) has been considered to be the most reliable indicator for FMD diagnosis. The current study extended the earlier designed recombinant 3AB3 protein-based indirect ELISA originally validated on bovine serum samples to testing serum samples of goat. The performance of the indirect ELISA was validated using internationally accepted PrioCHECK® FMDV NS kit. The overall diagnostic sensitivity (DSn) of the indirect ELISA was estimated to be 95.52% (619/648), while the diagnostic specificity (DSp) on naïve and vaccinated animals varied at 98.06% (557/568) and 94.15% (435/462), respectively. In India, where FMD is prevalent and the goat population is so high, this 'in-house' optimized assay can be considered to be an adjunct in sero-epidemiological investigation of FMD in goat.

Design and Development of an Antigen Test for SARS-CoV-2 Nucleocapsid Protein to Validate the Viral Quality Assurance Panels.

The continuing mutability of the SARS-CoV-2 virus can result in failures of diagnostic assays. To address this, we describe a generalizable bioinformatics-to-biology pipeline developed for the calibration and quality assurance of inactivated SARS-CoV-2 variant panels provided to Radical Acceleration of Diagnostics programs (RADx)-radical program awardees. A heuristic genetic analysis based on variant-defining mutations demonstrated the lowest genetic variance in the Nucleocapsid protein (Np)-C-terminal domain (CTD) across all SARS-CoV-2 variants. We then employed the Shannon entropy method on (Np) sequences collected from the major variants, verifying the CTD with lower entropy (less prone to mutations) than other Np regions. Polyclonal and monoclonal antibodies were raised against this target CTD antigen and used to develop an Enzyme-linked immunoassay (ELISA) test for SARS-CoV-2. Blinded Viral Quality Assurance (VQA) panels comprised of UV-inactivated SARS-CoV-2 variants (XBB.1.5, BF.7, BA.1, B.1.617.2, and WA1) and distractor respiratory viruses (CoV 229E, CoV OC43, RSV A2, RSV B, IAV H1N1, and IBV) were assembled by the RADx-rad Diagnostics core and tested using the ELISA described here. The assay tested positive for all variants with high sensitivity (limit of detection: 1.72-8.78 ng/mL) and negative for the distractor virus panel. Epitope mapping for the monoclonal antibodies identified a 20 amino acid antigenic peptide on the Np-CTD that an in-silico program also predicted for the highest antigenicity. This work provides a template for a bioinformatics pipeline to select genetic regions with a low propensity for mutation (low Shannon entropy) to develop robust 'pan-variant' antigen-based assays for viruses prone to high mutational rates.

Extracellular Vesicle Tissue Factor Activity Assay.

Tissue factor (TF) is a transmembrane receptor for factor (F) VII and FVIIa. The TF/FVIIa complex initiates the coagulation cascade by activating both FIX and FX. TF is released from cells into the circulation in the form of extracellular vesicles (EVs). The level of TF-positive (+) EVs is increased in various diseases, including cancer, bacterial and viral infections, and cirrhosis, and is associated with thrombosis, disseminated intravascular coagulation, disease severity, and mortality. There are two ways to measure TF+ EVs in plasma: antigen- and activity-based assays. Data indicates that activity-based assays have higher sensitivity and specificity than antigen-based assays. This paper describes our in-house EVTF activity assay based on a two-stage FXa generation assay. FVIIa, FX, and calcium are added to the TF+ EV-containing samples to generate FXa in the presence and absence of anti-TF antibody to distinguish TF-dependent FXa generation from TF-independent FXa generation. A chromogenic substrate cleaved by FXa is used to determine the FXa level, while a standard curve generated with a relipidated recombinant TF is used for the determination of the TF concentration. This in-house EVTF activity assay has higher sensitivity and specificity than a commercial TF activity assay.

Antigen-Based Diagnosis of Human Giardiasis: A Systematic Review and Meta-Analysis.

We aimed to present a systematic review and meta-analysis of studies that used antigen-based assays for the diagnosis of human giardiasis. All the related published literature cited within PubMed, ISI web of science, Google Scholar, Embase, and Scopus, were searched up to December 2021. The search terms, both as MeSH terms and text words, were "Giardia", "Giardia lamblia", "Giardia intestinalis", "giardiasis", combined with "diagnosis", "antigen detection", serodiagnosis, or serological diagnosis. The required data was extracted from the papers. Pooled estimates of sensitivity and specificity were obtained and forest plots and summary receiver operating characteristics (SROC) plots were used to calculate sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The search of databases found 1683 papers, of which 46 articles fulfilled our eligibility criteria. The sensitivity of antigen-based methods for the diagnosis of human giardiasis ranged from 45% (95% CI: 31-59%) to 100% (95% CI: 100-100%) and the pooled estimate of sensitivity was 92% (95% CI: 90-93%). The pooled estimated specificity was 97% (95% CI: 96-98%), ranged from 81% (95% CI: 68-89%) to 100% (95% CI: 98-100%). The summary estimate of PPV and NPV were 92 % (95% CI: 90-93%) and 97% (95% CI: 96-98%) respectively. Comparing the performance of the antigen detection assays by region revealed a significant difference in the assay's performances in different regions of the world. The antigen-based detection methods have acceptable and satisfactory performance in the diagnosis of human giardiasis. The task ahead is to identify more specific target antigens and design simpler, cheaper, and more sensitive methods for the diagnosis of this common worldwide-distributed parasitic infection.

Diagnosis of protozoa diarrhoea in Campylobacter patients increases markedly with molecular techniques.

Cryptosporidium and Giardia are major causes of diarrhoea globally, and two of the most notified infectious diseases in New Zealand. Diagnosis requires laboratory confirmation carried out mostly via antigen or microscopy-based techniques. However, these methods are increasingly being superseded by molecular techniques. Here we investigate the level of protozoa detection by molecular methods in campylobacteriosis cases missed through antigen-based assays and investigate different molecular testing protocols. We report findings from two observational studies; the first among 111 people during a Campylobacter outbreak and the second during normal surveillance activities among 158 people presenting with diarrhoea and a positive Campylobacter test, but negative Cryptosporidium and Giardia antigen-based test results. The molecular methods used for comparison were in-house end-point PCR tests targeting the gp60 gene for Cryptosporidium and gdh gene for Giardia. DNA extraction was performed with and without bead-beating and comparisons with commercial real-time quantitative (qPCR) were made using clinical Cryptosporidium positive sample dilutions down to 10-5. The Cryptosporidium prevalence was 9% (95% CI: 3-15; 10/111) and Giardia prevalence 21% (95% CI: 12-29; 23/111) in the 111 Campylobacter outbreak patients. The Cryptosporidium prevalence was 40% (95% CI: 32-48; 62/158) and Giardia prevalence 1.3% (95% CI: 0.2-4.5; 2/158) in the 158 routine surveillance samples. Sequencing identified Cryptosporidium hominis, C. parvum, and Giardia intestinalis assemblages A and B. We found no statistical difference in positive test results between samples using end-point PCR with or without bead-beating prior to DNA extraction, or between the in-house end-point PCR and qPCR. The qPCR Ct value was 36 (95% CI: 35-37) for 1 oocyst, suggesting a high limit of detection. In conclusion in surveillance and outbreak situations we found diagnostic serology testing underdiagnoses Cryptosporidium and Giardia coinfections in Campylobacter patients, suggesting the impact of protozoa infections may be underestimated through underdiagnosis using antigen-based assays.

Update of European Society of Clinical Microbiology and Infectious Diseases coronavirus disease 2019 guidelines: diagnostic testing for severe acute respiratory syndrome coronavirus 2

ScopeSince the onset of COVID-19, several assays have been deployed for the diagnosis of SARS-CoV-2. The European Society of Clinical Microbiology and Infectious Diseases (ESCMID) published the first set of guidelines on SARS-CoV-2 in vitro diagnosis in February 2022. Because the COVID-19 landscape is rapidly evolving, the relevant ESCMID guidelines panel releases an update of the previously published recommendations on diagnostic testing for SARS-CoV-2. This update aims to delineate the best diagnostic approach for SARS-CoV-2 in different populations based on current evidence. MethodsAn ESCMID COVID-19 guidelines task force was established by the ESCMID Executive Committee. A small group was established, half appointed by the chair, and the remaining selected with an open call. The panel met virtually once a week. For all decisions, a simple majority vote was used. A list of clinical questions using the population, intervention, comparison, and outcome (PICO) format was developed at the beginning of the process. For each PICO, 2 panel members performed a literature search focusing on systematic reviews with a third panellist involved in case of inconsistent results. The panel reassessed the PICOs previously defined as priority in the first set of guidelines and decided to address 49 PICO questions, because 6 of them were discarded as outdated/non-clinically relevant. The ‘Grading of Recommendations Assessment, Development and Evaluation (GRADE)-adoption, adaptation, and de novo development of recommendations (ADOLOPMENT)’ evidence-to-decision framework was used to produce the guidelines. Questions addressed by the guidelines and recommendationsAfter literature search, we updated 16 PICO questions; these PICOs address the use of antigen-based assays among symptomatic and asymptomatic patients with different ages, COVID-19 severity status or risk for severe COVID-19, time since the onset of symptoms/contact with an infectious case, and finally, types of biomaterials used.

Potential diagnostic application of the baculovirus-expressed recombinant truncated nucleocapsid protein of peste des petits ruminants virus in ELISA

The study describes the expression of recombinant truncated nucleocapsid protein (NP) of peste des petits ruminants (PPR) virus in the baculovirus system (PPRV-rBNP) and its potential application as a diagnostic antigen in ELISA for diagnosis of PPR in sheep and goats. The PPRV N-terminal immunogenic region (1–266 aa) of the NP coding sequence was amplified and cloned into the pFastBac HT A vector. The PPRV-rBNP with a molecular weight of ∼30 kDa was expressed in an insect cell system using generated recombinant baculovirus through Bac-to-Bac® Baculovirus Expression System. The crude PPRV-rBNP or Ni-NTA affinity-purified NP was characterized by SDS-PAGE and immunoblot using standard PPRV-specific sera. The PPRV-rBNP reacted well with PPRV anti-N specific monoclonal and polyclonal antibodies and PPRV-specific antiserum, suggesting that the expressed PPRV-rBNP is in its native form. The crude PPRV-rBNP as a diagnostic antigen was evaluated either as a coating antigen or standard positive control antigen in the Avidin-Biotin ELISA using the known standard panel reagents. The results showed that the expressed PPRV-rBNP can be an alternative diagnostic antigen to E. coli expressed recombinant PPRV-NPN and the utility of PPRV-rBNP avoids the need to use live PPRV antigen in the diagnostic ELISA. Hence, this allows scope in the future for large-scale field application of the recombinant antigen-based assays for diagnosis/surveillance and monitoring of PPR at the eradication as well as post-eradication phases in endemic countries or PPR non-endemic countries.

A Nationwide Survey on Detection of Chlamydia trachomatis in Health Facilities in China.

This study evaluated the clinical laboratory capacity for the diagnosis of Chlamydia trachomatis (CT) in China to provide recommendations to improve the diagnostic capacity and quality of this clinically important sexually transmitted disease. An electronic questionnaire-based cross-sectional, survey study was conducted by the National Center for STD Control among different types of healthcare facilities in China from July to December 2021. The surveyed laboratory facilities were located in 332 cities in 31 provinces in China. A total of 4640 records from clinical laboratories were included in the data set for the final analysis. Less than half of the laboratories (41.6% [1931 of 4640]) performed the CT diagnostic test; of these, 721 laboratories (15.5% [721 of 4640]) carried out nucleic acid amplification test (NAAT) methods, and 1318 laboratories (28.4% [1318 of 4640]) performed antigen-based immunochromatographic assays. Most laboratories were equipped with biological safety cabinets (93.7% [4348 of 4640]), 49.2% (2283 of 4640) were equipped with fully automated nucleic acid extractors, and 55.2% (2560 of 4640) were equipped with polymerase chain reaction amplification instruments. The laboratories from Southern China or third-class hospitals (i.e., the highest rated hospitals) had the highest proportion using NAATs to diagnose CT among the surveyed health facilities. Advancing laboratories to use NAAT to detect CT should be phased step-by-step by different areas and levels of hospitals according to the current situation.

Monkeypox infection elicits strong antibody and B cell response against A35R and H3L antigens

Monkeypox virus (MPXV) resides in two forms; mature and enveloped, and depending on it, distinct proteins are displayed on the viral surface. Here, we expressed two MPXV antigens from the mature, and one from the enveloped form, and tested their reactivity to sera of 11 MPXV recoverees while comparing to sera from recently and past vaccinated individuals. 8 out of 11 recoverees exhibited detectable neutralization levels against Vaccinia Lister. Sera from all recoverees bound strongly to A35R and H3L antigens. Moreover, the responses to A35R were significantly higher within the recoverees compared to both recently and past vaccinated donors. Lastly, A35R- and H3L-specific IgG+ B cells ranging from 0.03-0.46% and 0.11-0.36%, respectively, were detected in all recoverees (A35R), and in 9 out of 11 recoverees (H3L). Therefore, A35R and H3L represent MPXV immune targets and could be used in a heat-inactivated serological ELISA for the identification of recent MPXV infection.

Utilizing Recombinant Reporter Henipaviruses to Conduct Antiviral Screening.

Spillovers of Nipah virus (NiV) from its pteropid bat reservoir into the human population continue to cause near-annual outbreaks of fatal encephalitis and respiratory disease in Bangladesh and India since its emergence in Malaysia over 20years ago. The current lack of effective antiviral therapeutics against NiV merits further testing of compound libraries against NiV using rapid quantitative antiviral assays. The development of recombinant henipaviruses expressing reporter fluorescence and/or luminescence proteins has facilitated the screening of such libraries. In this chapter, we provide a basic protocol for both types of reporter viruses. Utilizing these live NiV-based reporter assays requires modest instrumentation and sidesteps the labor-intensive steps associated with traditional cytopathic effect or viral antigen-based assays.

Expression and Evaluation of Wb-SXP-1 and Wb-123 Recombinant Antigens as Potential Diagnostic Biomarkers for Lymphatic Filariasis

Lymphatic filariasis (LF) remains a public health concern as it can cause permanent morbidity and disability to those infected. While the global elimination of LF in these endemic areas is ongoing through mass drug administration, there is the need to develop diagnostic tools that would be utilized to track the progress of total global eradication as well as perform surveillance for the recurrence of lymphatic filariasis transmission. Currently, approved LF diagnosis tools are faced with lack of specificity, low sensitivity, and periodicity dependence. Recombinant filarial antigen-based assays can address these drawbacks and offer practical instruments for LF diagnosis and surveillance. This present study, evaluated rWb-SXP-1 and rWb-123 antigens as potential diagnostic biomarker tools for Wuchereria banchrofti in human sera using microspheres-based multiplex serological assay. Based on statistical analysis using XLSTAT 2019 (Addinsoft) on data generated from multiplex technology assay, generated ROC curves for both rWb-SXP-1 and rWb-123 demonstrated 87.1% sensitivity to Wuchereria banchrofti human sera with rWb-SXP-1 antigens having the highest specificity of 96%. Indication that rWb-SXP-1 and rWb-123 antigens are capable of detecting immunoglobulin G4 (IgG4) antibodies in human sera synthesized specifically against W. banchrofti infections. Therefore, rWb-SXP-1 and rWb-123 antigens can be utilized to detect W. banchrofti infections by antibody profiling with excellent diagnostic sensitivity and specificity using microsphere-based multiplex serological tests. This method can be particularly practical for screening a large number of sera samples and/or for quick, extensive field-testing due to the high-throughput and quick formats applied.

1383. Necrotizing Fasciitis and Streptococcus pyogenes (GAS) Infection Seasonality and Risk Factors for Infection— a Multicenter Research Network Study

Abstract Background Streptococcus pyogenes causes acute pharyngitis and type II necrotizing fasciitis. Seasonal variations in the incidence of S. pyogenes infections are not robustly characterized. We aim to identify seasonal variations and risk factors of S. pyogenes infections and all causes of necrotizing fasciitis. Methods From 2010 to 2019, we identified all infectious adult cases of S. pyogenes using ICD-10 diagnoses and lab results (PCR and antigen-based assays) and cases of necrotizing fasciitis using ICD-10 diagnoses within a federated research network. We extracted seasonal (quarterly) incidence rates. We used an autoregressive integrated moving average (ARIMA) model to assess the seasonality of the cases (6-month intervals). Demographic characteristics and 3-month outcomes of S. pyogenes pharyngitis were compared to a cohort of patients with acute upper respiratory illnesses excluding S. pyogenes. Results We identified 238,088 cases of S. pyogenes pharyngitis and 26,931 cases of necrotizing fasciitis in adults. S. pyogenes infection average incidence was higher during the winter than the summer: 0.34 vs. 0.20 cases per 1,000 patients. Necrotizing fasciitis diagnoses were highest during the summer months (average 0.026 per 1000 patients). There was a significant ARIMA seasonal variation in the time series analysis for S. pyogenes infections (p=0.006) (figure 1). However, necrotizing fasciitis was not significant (p=0. 0.79) (Figure 2). Compared to adults with acute respiratory infections other than S. pyogenes, adults with S. pyogenes pharyngitis were more likely to be younger (25.8 ± 14.9 vs. 45.4 ± 19.9 years old, p< 0.0001) and of Hispanic or Latino ethnicity (11% vs. 8%, p < 0.0001). For age-matched outcomes, adults with S. pyogenes pharyngitis had lower rates of hospitalization (0.888% vs. 1.714%, p< 0.0001) and mortality (0.114% vs. 0.174%, p< 0.0001) at three months relative to adults with acute respiratory infections other than S. pyogenes. Figure 1.Seasonal variation of adults with established ICD code or testing for Streptococcus pyogenes (GAS) infectionsFigure 2.Seasonal variation of adults with established ICD code for necrotizing fasciitis Conclusion Peaks in S. pyogenes infections are more likely to occur in the winter months, although spring and fall seasons can display variably high rates of S. pyogenes year over year. Necrotizing fasciitis of any microbiological did not show a significant seasonal variation. Disclosures All Authors: No reported disclosures.

Viral Load-based Evaluation of the Sensitivity of Antigen-based Rapid Detection Assay for SARS-CoV-2

Viral Load-based Evaluation of the Sensitivity of Antigen-based Rapid Detection Assay for SARS-CoV-2

An Illustrative Model of Economically Utilizing Point of Care and Reverse Transcriptase Polymerase Chain Reaction Testing to Reduce Costs and Help Balance Supply and Demand

Abstract Introduction/Objective Both point of care (POC) antigen-based assays and reverse transcriptase polymerase chain reaction (RT-PCR) laboratory-based assays exist for diagnosis of COVID-19. COVID-19 RT-PCR tests are the gold standard based on their higher sensitivity. However, in this era of inflation and supply chain disruptions, the use of only RT-PCR tests would be more expensive and may be vulnerable to reagent supply chain shortages. Triage of resources of both test types is important in ensuring access and equity in testing distribution as well as ensuring economical RT-PCR testing access for sicker patients. A comparison of costs was explored by contrasting a model of cost effectively reserving RT-PCR tests for those with significant disease requiring hospitalization and using POC tests for those with mild disease (defined as at most mild symptoms not requiring hospitalization) versus universal provision of RT-PCR to all. Methods/Case Report Cost information from the regional Veteran Affairs Medical Center (VAMC), specifically for the Alinity-m (RT-PCR) and BinaxNOW (POC) by Abbott (Chicago IL), and from the community (median COVID-19 RT- PCR pricing obtained from the Peterson KFF Health System Tracker) were included. Based on this cost information, a model of 100 tested COVID-19 suspected patients was created. A percentage of mild disease not requiring hospitalization (81%) was utilized based on literature from early in the COVID-19 pandemic. Results (if a Case Study enter NA) The cost per test at VAMC factored in was $28.34 (Alinity-m) and $5 (BinaxNOW). The median community RT-PCR and POC price factored in was $148 and $25, respectively. From the model of 100 COVID-19 with a mild disease percentage of 81%, the total testing cost of including POC assays for mild cases vs. universal RT-PCR assays would be $943.46 vs. $2,834 at the VAMC and similarly $4,837 vs. $14,800 in the community for a 67% reduction in costs at both settings. Conclusion A model of testing including both POC and RT-PCR testing may be economical and critically help ensure triage of resources in this era of inflation and supply chain shortages.

A Cost Comparison Modeling Analysis for Healthcare Access and Equity: Cost of COVID-19 Test Screening by Point of Care (POC) Versus Rapid Reverse Transcriptase Polymerase Chain Reaction (RT-PCR)

Abstract Introduction/Objective Testing platforms to detect COVID-19 infection include antigen-based point of care (POC) testing and reverse transcriptase polymerase chain reaction (RT-PCR) to detect viral ribonucleic acid (RNA). RT-PCR, generally considered the gold standard for diagnosis of COVID-19 infection, is capable of being used for mass population screening but may be more expensive. Alternatively, antigen based POC testing can not only produce results rapidly (15 minutes on some assays) but also may be less expensive. This reduced cost provides an argument that it may be economical to use POC testing for mass surveillance in asymptomatic or mildly symptomatic patients while reserving the more expensive gold standard RT-PCR testing for sensitivity for hospitalized and more significantly at risk or ill patients. However, cost modeling between the two modalities for mass screening is sparse, particularly for studies including the Veteran Affairs system that may have different costs from the general community retail pricing. Methods/Case Report Cost information reviewed at the regional VAMC included costs for the Alinity-m SARS-CoV-2 RT-PCR assay (Abbott, Chicago IL), the Cepheid Xpert-Xpress-CoV-2-Flu-RSV-Plus RT-PCR assay (Cepheid, Sunnyvale CA), and the BinaxNOW POC antigen-based assay (Abbott, Chicago IL). The median cost of RT-PCR testing in the community was obtained from the Peterson-KFF Health System Tracker. Publicly available quoted prices for selected POC assays in the community were reviewed for comparison. Results (if a Case Study enter NA) Test costs at the regional VAMC was $28.34 (Alinity-m), $68 (Cepheid), and $5 (BinaxNOW). These costs did not include technologist time or standard laboratory consumables that would be part of the standard overhead costs of running the laboratory. The median community RT-PCR assay price per the Peterson- KFF Health System Tracker was $148, and the Abbott’s quoted BinaxNOW price was $25. POC testing cost was 82% less than the cheapest RT-PCR assay available at the regional VAMC. This trend in being significantly cheaper holds true in the community whereby BinaxNOW testing was 83% cheaper. Publicly available quoted prices from other selected POC assays confirm this trend. Conclusion For a mass surveillance screening program, the use of POC assays would result in significant cost savings.