complex sample types. However, in order to remove potentially interfering particles including endogenous antibodies such as heterophilic antibodies (HA) (e.g. HAMA) and rheumatoid factor (RF), it is important to incorporate immunoassay blockers into the assay design. Double mouse monoclonal assays such as those for rapid respiratory antigen tests, are specifically prone to HAMA and RF interference and require a specialized blocker to ensure the assay’s accuracy. Meridian manufactures in multikilogram scales of passive blockers such as Mouse IgG and animal serums as well a proprietary active blocker, TRU Block ™ , which contains specific binders directed against all types of heterophilic interference including HAMA and RF. Once bound to the interfering antibodies, TRU Block ™ prevents further binding of HA to other assay components through steric hindrance. Active blockers can typically be used in lower concentrations than passive blocking reagents, which minimizes the reduction in assay signal commonly associated with passive blockers. Overall, immunoassay multiplexing calls for
carefully chosen reagents that allow the antibodies to work together to produce an accurate, meaningful result. Interfering factors must be minimized to prevent a false positive or a false negative error. Conclusion Multiplex assays that combine the detection of several targets at once have several advantages including their high-throughput potential, ability to provide more results per sample, and lower reagent consumption (i.e., miniaturization). All of these benefits translate to a lower price-per-data point compared to traditional singleplex assays. With the reopening of most countries and relaxing of social distancing and mask wearing strategies, the circulation of influenza, COVID-19 and RSV will rebound and likely exceed normal levels for some time. Given many respiratory disease symptoms overlap, it is not possible to reliably differentiate an infection with one of these viruses on clinical grounds alone. Multiplex testing addresses the higher demand for screening between infections due to SARS-CoV-2, influenza A/B and RSV during what is expected to be higher than normal flu season. Molecular multiplex
testing offers high sensitivity and specificity, detecting infected individuals early in the course of the disease and enabling adequate time for the appropriate treatment. However, molecular tests require sophisticated equipment, experienced technicians and generally have a turn-around time of 1-5 days. In contrast, immunoassay rapid antigen assays are easy to use and can provide a result in less than 10-15 minutes, but they are considered to have a lower performance and are less sensitive. Each testing type, molecular or immunoassay, has advantages and disadvantages and they can be used independently from one another or in a testing algorithm that leverages the advantages of both. By employing multiplexing assays for the flu, COVID and RSV, patients will be able to be treated faster and more effectively, regardless of the testing type employed.
1. Centers for Disease Control and Prevention. Rapid Diagnostic Testing for Influenza: Information for Clinical Laboratory Directors. https://www.cdc.gov/flu/professionals/diagnosis/rapidlab.htm. Accessed October 26, 2021 2. Jong, Jong-Yoon. (February 2013). Multiplex molecular diagnostics: shifting the paradigm. Medica Laboratory Observer. Retrieved from https://www.mlo-online.com/home/article/13005002/ multiplex-molecular-diagnostics-shifting-the-paradigm 3. Centers for Disease Control and Prevention. RSV in Infants and Young Children https://www.cdc. gov/rsv/high-risk/infants-young-children.html. Accessed October 26, 2021. 4. Meng, J., Stobart, C. C., Hotard, A. L. & Moore, M. L. An overview of respiratory syncytial virus. PLoS Pathog. 10, e1004016 (2014).
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