A fluorescent microspheres-based microfluidic test system for the detection of immunoglobulin G to SARS-CoV-2

Background: The pandemic of the new coronavirus infection, COVID-19, is currently ongoing in the world. Over the years, the pathogen, SARS-CoV-2, has undergone a series of mutational genome changes, which has led to the spread of various genetic variants of the virus. Meanwhile, the methods used to diagnose SARS-CoV-2, to establish the disease stage and to assess the immunity, are nonspecific to SARS-CoV-2 variants and time-consumable. Thus, the development of new methods for diagnosing COVID-19, as well as their implementation in practice, is currently an important direction. In particular, application of systems based on chemically modified fluorescent microspheres (with a multiplex assay for target protein molecules) opens great opportunities. Aim: development of a microfluidic diagnostic test system based on fluorescent microspheres for the specific detection of immunoglobulins G (IgG) to SARS-CoV-2. Methods: A collection of human serum samples was characterized using enzyme-linked immunosorbent assay (ELISA) and commercially available reagent kits. IgG to SARS-CoV-2 in the human serum were detected by the developed immunofluorescent method using microspheres containing the chemically immobilized RBD fragment of the SARS-CoV-2 (Kappa variant) viral S-protein. Results: The level of IgG in the blood serum of recovered volunteers was 9-300 times higher than that in apparently healthy volunteers, according to ELISA (p0.001). Conjugates of fluorescent microspheres with the RBD-fragment of the S-protein, capable of specifically binding IgG from the blood serum, have been obtained. The immune complexes formation was confirmed by the fluorescence microscopy data; the fluorescence intensity of secondary antibodies in the immune complexes formed on the surface of microspheres was proportional to the content of IgG (r 0.963). The test system had a good predictive value (AUC 70.3%). Conclusion: A test system has been developed, based on fluorescent microspheres containing the immobilized RBD fragment of the SARS-CoV-2 S-protein, for the immunofluorescent detection of IgG in the human blood serum. When testing the system on samples with different levels of IgG to SARS-CoV-2, its prognostic value was shown. The obtained results allow us to present the test system as a method to assess the level of immunoglobulins to SARS-CoV-2 in the human blood serum for the implementation in clinical practice. The test system can also be integrated into various microfluidic systems to create chips and devices for the point-of-care diagnostics

International train the trainer neonatal antibiotic stewardship program for South African pharmacists

Hospital-acquired antimicrobial-resistant infections are a leading cause of neonatal mortality in South African (SA) neonatal intensive care units (NICU). There is an urgent need for NICU Antibiotic Stewardship Programs (ASP). We describe the development of an international Train-the-Trainer (TTT) NICU-ASP mentoring program for SA pharmacists. A partnership between the South Africa Antimicrobial Stewardship in 2019. A baseline assessment of four SA NICUs was done to guide the development of a TTT NICU-ASP mentoring of SA pharmacists utilizing the existing workforce. The program included bilateral site visits. Pre-post surveys were used to assess SA mentee's NICU experiences, barriers to clinical pharmacy services and confidence to train additional pharmacists in NICU ASP. Four mentees from private (n = 1) and public hospitals (n = 3) completed a 2-week TTT NICU-ASP in the US that included; education, patient care rounds, role-playing, peer-to-peer sessions and behavioral interventions followed by ongoing support and mentoring by SAASP mentors. None of the hospitals had pharmacists participating in daily patient care rounds or had multidisciplinary NICU-ASPs due to lack of NICU trained pharmacists and dedicated time for ASP. Post surveys showed improved confidence to train additional pharmacists in NICU-ASP. Subsequently, these SA mentees provided NICU-ASP education to over 700 health care professionals and trained six additional pharmacists in NICU-ASP. Mentors and mentees developed a comprehensive NICU ASP toolkit for ongoing training of additional pharmacists. A new research collaboration between TTT NICUASP mentors, mentees and physician members of the South Africa National Neonatal Sepsis Task Force has formed and the first national NICU-ASP study is underway in 12 hospitals. Shared leadership between U.S. and SA mentors led to developing a TTT NICU-ASP for pharmacists tailored to existing resources and local needs.Merck; Pfizer; Bill and Melinda Gates Foundation grant.http://wileyonlinelibrary.com/journal/jac5am2022Pharmacolog

A global point prevalence survey of antimicrobial use in neonatal intensive care units: The no-more-antibiotics and resistance (NO-MAS-R) study

Background: Global assessment of antimicrobial agents prescribed to infants in the neonatal intensive care unit (NICU) may inform antimicrobial stewardship efforts. Methods: We conducted a one-day global point prevalence study of all antimicrobials provided to NICU infants. Demographic, clinical, and microbiologic data were obtained including NICU level, census, birth weight, gestational/chronologic age, diagnoses, antimicrobial therapy (reason for use; length of therapy), antimicrobial stewardship program (ASP), and 30-day in-hospital mortality. Findings: On July 1, 2019, 26 of infants (580/2,265; range, 0�100; median gestational age, 33 weeks; median birth weight, 1800 g) in 84 NICUs (51, high-income; 33, low-to-middle income) from 29 countries (14, high-income; 15, low-to-middle income) in five continents received �1 antimicrobial agent (92, antibacterial; 19, antifungal; 4, antiviral). The most common reasons for antibiotic therapy were �rule-out� sepsis (32) and �culture-negative� sepsis (16) with ampicillin (40), gentamicin (35), amikacin (19), vancomycin (15), and meropenem (9) used most frequently. For definitive treatment of presumed/confirmed infection, vancomycin (26), amikacin (20), and meropenem (16) were the most prescribed agents. Length of therapy for culture-positive and �culture-negative� infections was 12 days (median; IQR, 8�14) and 7 days (median; IQR, 5�10), respectively. Mortality was 6 (42, infection-related). An NICU ASP was associated with lower rate of antibiotic utilization (p = 0·02). Interpretation: Global NICU antibiotic use was frequent and prolonged regardless of culture results. NICU-specific ASPs were associated with lower antibiotic utilization rates, suggesting the need for their implementation worldwide. Funding: Merck & Co.; The Ohio State University College of Medicine Barnes Medical Student Research Scholarship © 2021 The Author