The Africa Working Group, under the auspices of the Global Laboratory Quality Initiative of the Association for Diagnostics & Laboratory Medicine (ADLM), conducted several educational workshops in Africa to promote laboratory quality in the region. The challenges faced in maintaining quality in clinical laboratories in low-resource countries may differ from region to region and country to country. To better understand current gaps and inform our engagement with partners in Africa, a needs assessment survey was distributed to 1427 ADLM members in Africa and further disseminated through local partners, resulting in 121 responses from participants across 8 countries in the region. Additionally, interviews were conducted with laboratory professionals from the countries surveyed. The surveys primarily focused on quality management, continuing education, budgeting, infrastructure and funding, supplies, and staffing issues. The data collected from the surveys and interviews strongly indicate that respondents face major challenges in all areas assessed, including lack of funding, lack of equipment, lack of quality control (QC) and external quality assessment (EQA) materials, gaps in quality management, lack of appropriate budgeting, staffing issues, and lack of accessible training/continuing education. Consistent with the primary focus of ADLM;s Global Quality Initiative, improved access to continuing education was identified as the primary need by the participants (58%). The assessment highlights that while a strong role for educational workshops in mitigating some of the challenges continues to exist, there is also an opportunity to broaden the scope of the working groups beyond education.

Laboratory diagnosis of syphilis can use either the usual or the reverse algorithm. In the usual algorithm, a reactive rapid plasma reagin (RPR) test is followed by a titer and measurement of treponemal antibodies. When using the reverse algorithm, patients are screened with a treponemal antibody test, and any reactive result is followed by an RPR. Treponemal antibody tests measure IgG and IgM against Treponema pallidum. These tests convert a number read by the instrument into a qualitative result. The objective of this study was to see if the number read by 2 treponemal antibody tests correlate with RPR titers. Fifty-one patients with reactive RPR tests were tested using the Roche Cobas instrument (which gives a cutoff index –COI result) and the Diasorin Liaison instrument (which gives an arbitrary unit –AU result). We correlated the RPR titer of each sample with the COI and AU. Then, retrospectively reviewed patient charts to define disease or potential false positive RPR results. There were 13 patients with RPR titers of 1:1. Of these only 1 had the diagnosis of syphilis. The average read in the Roche was 105 (range 28.3-252) while the Diasorin was 31.6 (range 8.4->70). The patient with the diagnosis of secondary syphilis had a COI of 158 and an AU of 47. There were 9 patients with RPR titers of 1:2. Of these 8 had diagnosis of current or treated syphilis. The average read in the Roche was 206 (18.8-357) while the Diasorin was 46 (9.4->70). Twenty seven of the 29 remining patients had diagnosis of current or treated syphilis. The 1:4 and 1:8 titers showed an increase in the average COI and AU for the Roche (214, 245 respectively) and Diasorin (51, 63) though with large overlap in the range. However, RPR titers above 1:8 did not show increase in the average results for Roche or Diasorin. Additionally, individual patient review did not necessarily correlate with titer. There appears to be a correlation of the average treponemal antibody instrument read for both the Roche and Diasorin platforms in patients with lower RPR titers. Though, the overlap in range likely precludes the use of the COI or AU read as a surrogate for RPR titers. The correlation for higher titers was not observed in our cohort of patients which could be due to the low numbers of patients in each category or the stage of the disease.

A sentinel event occurred one year after the opening of a new 100-bed cancer hospital. During surgery, a type B patient required multiple units of red blood cells (RBCs) and the operating room (OR) team started retrieving products from a remote refrigerated blood storage system (RRBSS) located near the OR. Compatible units were retrieved from the RRBSS, but depletion of these units did not allow the OR team to retrieve plasma products also stored in the RRBSS. After talking with the blood bank, units were issued but the medical laboratory scientists were trying to provide type-specific products rather than treating this as a massive transfusion protocol (MTP) activation because an MTP was not explicitly ordered. Here we describe a quality improvement process for MTP in a new hospital. Root cause analyses (RCA) regarding the processes in the OR and the laboratory were conducted. We reviewed the MTP process used in a nearby trauma hospital and what other hospitals did with their RRBSS regarding MTPs. We talked with attending surgeons, anesthesia and nursing in our hospital. Then, the OR team and the laboratory created an escalation plan, educational materials for the OR, and created a protocol for the laboratory. We conducted a simulation with all personnel involved and debrief with perioperative staff following MTP activations. The OR RCA uncovered gaps in knowledge for OR staff that prevented retrieval of the required products from the RRBSS. Also, anesthesia personnel did not know how to order an MTP on the computer system or how to activate it by calling the laboratory. The laboratory RCA uncovered that it took longer than expected to prepare MTP packages because newer technologists were not familiar with the protocol and partly because of where the products were stored. The following actions were implemented: education was provided to OR staff on how to activate a massive transfusion protocol electronically and verbally. We distributed “cheat sheets” about the appropriate use and activation of MTPs to the OR. The laboratory educated technologists on MTPs and implemented a drawer in one of the refrigerators that contains RBCs and thawed fresh frozen plasma units needed for the first MTP cooler. The laboratory process implemented states that as soon as the first cooler is sent to the OR, the laboratory prepares the next cooler. The mock simulation took place two weeks after the sentinel event. After implementing the MTP process improvement, 3 subsequent massive transfusion events have occurred in the hospital with minimal delays in providing necessary products.

OBJECTIVE:Clinical pathology curriculum assessment at our institution suggested improvements in coagulation, histocompatibility, and immunology. We created a 4-week rotation inclusive of these fields, and we describe here the curriculum and experience residents have had. METHODS:Didactic lectures, assigned reading, time at the bench, online modules, and case study discussions were implemented. There were 2 weeks of morning activities dedicated to histocompatibility, while the other 2 weeks were dedicated to immunology. Afternoons were dedicated to special coagulation. Resident surveys and medical knowledge assessments were completed at the beginning and end of the rotation. RESULTS:We included in this analysis 22 residents. The average knowledge quiz score before the rotation was 15.7 (48% [range, 8-23]) correct answers, while the average score after the rotation was 25 (76% [range, 19-29]), corresponding to a 28% increase in scores. On a Likert scale of 1 to 5, 88% of trainees ranked the rotation at a 4. After the rotation, residents felt more confident interpreting studies for the 3 subspecialties, particularly for special coagulation. CONCLUSIONS:Residents liked the rotation, and their evaluations demonstrate an increase in medical knowledge after the rotation. The training described here can be adapted to teach other compatible clinical pathology subspecialties.

Hemostatic alterations are common in patients with myeloproliferative neoplasia (MPN), which may increase the risk of both bleeding and thrombosis. Specifically, acquired von Willebrand syndrome (AVWS) is a known complication of MPN. On the other hand, aspirin is a frequently used treatment for the prevention of thrombotic events in patients with MPN. An increased bleeding risk associated with aspirin use has been found in patients harboring CALR mutations or platelet counts >1,000x109/L. The combination of platelet count >1,000x109/L, leukocytosis and AVWS has also been identified as a biological risk factor for bleeding. This retrospective study aimed to elucidate the clinical utility of von Willebrand studies in treatment decisions regarding prophylactic aspirin for MPN patients within our hospital system. We queried our electronic health record for patients with the diagnosis of MPN and von Willebrand disease (VWD) in the past 25 years. Data collected included patient demographic information, bleeding and treatment history, relevant laboratory data (e.g. platelet counts, VWD results), and results of the VWD panel testing (including factor VIII, VW antigen, and ristocetin/VW activity). Twenty patients were included in this study. All patients (100%) demonstrated a type 2-like AVWS, with activity to antigen ratio of < 0.7. Sixteen patients (80%) had essential thrombocythemia (ET), with the two most common mutations being CALR (45%) and JAK2 (30%). Eleven patients (55%) were on aspirin; among these, 65% had bleeding complications. There was no significant difference in VWD test parameters (i.e. factor VIII, VW antigen, VW activity, and activity/antigen ratio) between those with and without bleeding events. Maximum platelet counts ranged from 343,000 – 4,391,000 per microliter, and there was no significant difference between those with and without bleeding histories. Furthermore, there was no correlation between maximum platelet counts and VW activity. No standardized criteria currently exist for the diagnosis or management of patients with MPN and AVWS. Here we aimed to determine any association between VW activity, platelet count, type of MPN, and type of mutation with bleeding risk in patients on aspirin treatment; however, no significant relationship was identified in the limited study cohort. Future investigation on this frequently cited complication of MPN deserves more attention.

Abstract Background Many academic institutions recognize contributions to individual departments by awarding faculty departmental awards for their achievement in service, teaching and scholarship. Before 2019, our department had teaching awards chosen by residents, but there were no awards given to the faculty by peers for academic accomplishments. The objective of this abstract is to present the impact of the newly implemented departmental awards on the promotion and academic advancement of our faculty. Design Based on the already established awards in other departments within the School of Medicine a committee of six faculty members proposed six annual awards: Educator Impact, Outstanding Service, Outstanding Citizenship, Mentoring, and Basic and Clinical Science Publications. The committee defined activities and eligibility for each award category (for example, faculty could not receive the same award within 3 years). The categories and eligibility criteria are distributed to all faculty (around 120) each year. The first call for nominations occurred in 2019. Assessment of the awards program included the number of nominators and nominees each year and the impact on recipients’ academic advancement and recognition. Results In 2019, 17 faculty nominated 16 other faculty for the six awards. These numbers grew steadily, and by 2023, 42 faculty nominated 28 other faculty for the awards. The awards have been included in the curriculum vitae of five faculty who have been promoted to either the Associate Professor or Professor level and six faculty inducted into our institution’s Clinical Distinctions program. During the five years this awards program has existed, two faculty received two awards in different categories, whereas the remaining 26 awardees each received one award. Anecdotes from faculty during this time have been overwhelmingly positive. Conclusion Based on the increased engagement of faculty nominating their peers and the overall impact of the awards on faculty promotion and clinical distinction recognition, the new departmental awards program has been very successful.

OBJECTIVES:Mpox is a viral disease caused by monkeypox, a highly contagious orthopoxvirus that resulted in a global outbreak beginning in spring 2022. Diagnosis is confirmed via polymerase chain reaction (PCR) testing of swabs from mucocutaneous lesions. Rare reports have documented the histologic changes of mpox lesions, but the cytologic features have not been described. We present the cytology findings of samples taken from swabs of mucocutaneous mpox lesions in 3 different patients.METHODS:The patients were all male, aged 55, 43, and 37 years, all with mpox confirmed by PCR testing. Swabs from chest (cases 1 and 2) and tongue (case 3) lesions were directly sampled and submitted in Aptima (case 1) or PreservCyt solution (cases 2 and 3). Liquid-based preps were prepared and stained using the Papanicolaou method. Specimens were assessed for viral cytopathic changes.RESULTS:All cases showed nuclear cytopathic changes (enlarged nuclei with open chromatin and prominent red nucleoli), 2 cases demonstrated multinucleated keratinocytes, and 1 case showed potential Guarnieri bodies. The chromatin margination and nuclear molding typical of herpesviruses was not appreciated.CONCLUSIONS:The cytopathic changes of monkeypox are not specific, but their recognition could prompt appropriate PCR testing. Monkeypox shows distinct cytologic changes compared with herpesviruses.

CONTEXT.—:Urinalysis instrument-specific dip strips offer physicians qualitative results for actionable analytes (protein, glucose, leukocyte esterase, nitrates, hemoglobin, and ketones). OBJECTIVE.—:To explain a strategy implemented to support clinical decision-making by providing urine quantification of protein, glucose, white blood cells (WBCs), and red blood cells because of urine strip shortages. DESIGN.—:During shortages, we implemented an automated algorithm that triggered sending urine samples to the automation line for quantification of protein and glucose and ensured that urine microscopy was performed to obtain WBC and red blood cell counts. The algorithm printed 2 labels so nursing staff would collect 2 specimens. We monitored the turnaround time from the specimen being received in the laboratory to result verification, ensured that the culture reflex order was triggered, and tracked complaints by physicians regarding not having usual urinalysis results. Prior to implementation, correlation between sample types for protein and glucose measurement was found acceptable. RESULTS.—:The algorithm was put in place twice during 2022. The turnaround time of urine microscopic study was identical to that obtained when the urinalysis was done with the strips; however, the quantification of glucose and protein took approximately 30 minutes more. Urine reflex cultures were triggered correctly with the algorithm, as they were derived entirely from a WBC count higher than 10 per high-power field. During the shortage period we had only 1 complaint, by a physician wanting to have results of nitrates. CONCLUSIONS.—:During urine strip shortages, we successfully implemented a diversion algorithm that provided actionable urinalysis analytes in a timely manner with minimal provider complaints.

A knowledge gap exists between apheresis medicine (AM) physicians and providers who request the service, presenting challenges when coordinating care. We investigated an educational intervention consisting of a 40-min in-person evidence-based lecture for neurology residents, neurology attending physicians, and nephrology fellows. Pre-/post-testing demonstrated substantially improved understanding of apheresis mechanics, indications, complications, and patient consent. We advocate for more educational sessions about AM for the non-apheresis providers to increase awareness about apheresis and foster collaborative interdisciplinary efforts.

BACKGROUND:Therapeutic plasma exchange (TPE) is the primary intervention for treating symptomatic hyperviscosity from hypergammaglobulinemia, yet its efficacy for treating hyperviscosity related to hyperfibrinogenemia is unclear. OBJECTIVES:Define the safety and efficacy of TPE for critically ill COVID-19 patients with elevated blood viscosity from hyperfibrinogenemia. METHODS:We performed a prospective randomized controlled trial in critically ill COVID-19 patients in a single US healthcare system. Patients with hyperfibrinogenemia (>800 mg/dL) or elevated plasma viscosity (2.3-3.5 centipoise [cP]) were randomized to receive TPE on 2 consecutive days or continued standard of care (SOC). RESULTS:Twenty participants were enrolled, with 10 receiving TPE and 10 receiving SOC alone. Mean (±SEM) plasma viscosity decreased significantly from 2.35 cP (±0.12) to 1.61 cP (±0.03) in the TPE group and was unchanged in the SOC group (2.47 cP [±0.11] to 2.47 cP [±0.15]). Mean fibrinogen decreased from 934.0 mg/dL (±25.1) to 359.1 mg/dL (±22.5) after TPE vs from 859.6 mg/dL (±57.6) to 807.3 mg/dL (±63.1) in SOC. There was no significant difference in 28-day all-cause mortality between groups, with 2 deaths in the TPE cohort and 5 deaths in the SOC cohort (P = .13). No serious safety events related to TPE were reported. TPE significantly decreased biomarkers of inflammation (erythrocyte sedimentation rate and C-reactive protein) and endothelial activation (von Willebrand factor and factor VIII) but not hemostatic activation (prothrombin fragment 1.2, thrombin-antithrombin complex, and fibrin monomer) or immunoglobulin (IgG and IgM) levels. CONCLUSION:TPE is safe and effective for normalizing elevated blood viscosity from hyperfibrinogenemia in COVID-19 patients. Additional studies are needed to determine the impact of TPE on overall patient outcomes, including in those with non-COVID-19 conditions associated with hyperfibrinogenemia.

OBJECTIVES:Since laboratory critical values reflect such an abnormal pathologic state that there is imminent danger to the patient, it is crucial to deliver the result upon initial call with an escalation process when the initial call cannot occur. In our 8-hospital system, one of the hospitals used the escalation procedure twice as frequently compared with the other hospitals. This work presents hospital-wide quality improvement processes that decreased escalation of critical value calls so as to reach the same proportion of escalated calls compared to other hospitals in the system. METHODS:The laboratory met weekly with leaders of different hospital areas and quality management; they presented the interventions they implemented, and the laboratory monitored their progress. RESULTS:Monitoring and reviewing with providers the importance of critical values decreased temporarily escalated calls from 25% to 18%. Having a dedicated phone to call critical values in each hospital area decreased the calls in a sustained fashion, which now fluctuate between 9% and 14%. Other interventions, including having a dedicated person receiving critical value results, did not decrease escalated critical value calls. CONCLUSIONS:Having a dedicated phone in each hospital area that receives the initial critical value call simplifies and standardizes the process.

Patients with cystic fibrosis (CF) exhibit pronounced respiratory damage and were initially considered among those at highest risk for serious harm from SARS-CoV-2 infection. Numerous clinical studies have subsequently reported that individuals with CF in North America and Europe-while susceptible to severe COVID-19-are often spared from the highest levels of virus-associated mortality. To understand features that might influence COVID-19 among patients with cystic fibrosis, we studied relationships between SARS-CoV-2 and the gene responsible for CF (i.e., the cystic fibrosis transmembrane conductance regulator, CFTR). In contrast to previous reports, we found no association between CFTR carrier status (mutation heterozygosity) and more severe COVID-19 clinical outcomes. We did observe an unexpected trend toward higher mortality among control individuals compared with silent carriers of the common F508del CFTR variant-a finding that will require further study. We next performed experiments to test the influence of homozygous CFTR deficiency on viral propagation and showed that SARS-CoV-2 production in primary airway cells was not altered by the absence of functional CFTR using two independent protocols. On the contrary, experiments performed in vitro strongly indicated that virus proliferation depended on features of the mucosal fluid layer known to be disrupted by absent CFTR in patients with CF, including both low pH and increased viscosity. These results point to the acidic, viscous, and mucus-obstructed airways in patients with cystic fibrosis as unfavorable for the establishment of coronaviral infection. Our findings provide new and important information concerning relationships between the CF clinical phenotype and severity of COVID-19.

Infections caused by members of the mycobacterium tuberculosis complex [MTC] and nontuberculous mycobacteria [NTM] can induce widespread morbidity and mortality in people. Mycobacterial infections cause both a delayed immune response, which limits rate of bacterial clearance, and formation of granulomas, which contain bacterial spread, but also contribute to lung damage, fibrosis, and morbidity. Granulomas also limit access of antibiotics to bacteria, which may facilitate development of resistance. Bacteria resistant to some or all antibiotics cause significant morbidity and mortality, and newly developed antibiotics readily engender resistance, highlighting the need for new therapeutic approaches. Imatinib mesylate, a cancer drug used to treat chronic myelogenous leukemia [CML] that targets Abl and related tyrosine kinases, is a possible host-directed therapeutic [HDT] for mycobacterial infections, including those causing TB. Here, we use the murine Mycobacterium marinum [Mm] infection model, which induces granulomatous tail lesions. Based on histological measurements, imatinib reduces both lesion size and inflammation of surrounding tissue. Transcriptomic analysis of tail lesions indicates that imatinib induces gene signatures indicative of immune activation and regulation at early time points post infection that resemble those seen at later ones, suggesting that imatinib accelerates but does not substantially alter anti-mycobacterial immune responses. Imatinib likewise induces signatures associated with cell death and promotes survival of bone marrow-derived macrophages [BMDMs] in culture following infection with Mm. Notably, the capacity of imatinib to limit formation and growth of granulomas in vivo and to promote survival of BMDMs in vitro depends upon caspase 8, a key regulator of cell survival and death. These data provide evidence for the utility of imatinib as an HDT for mycobacterial infections in accelerating and regulating immune responses, and limiting pathology associated with granulomas, which may mitigate post-treatment morbidity.

Pathologists are an integral part of One Health as they are a critical component of the multidisciplinary team that diagnoses zoonotic diseases and discovers emerging pathogens. Both human and veterinary pathologists are uniquely positioned to identify clusters or trends in patient populations that can be caused by an infectious agent and preface emerging outbreaks. The repository of tissue samples available to pathologists is an invaluable resource that can be used to investigate a variety of pathogens. One Health is an encompassing approach that focuses on optimizing the health of humans, animals (domesticated and sylvatic), and the ecosystem, including plants, water, and vectors. In this integrated and balanced approach, multiple disciplines and sectors from local and global communities work together to promote overall well-being of the 3 components and address threats such as emerging infectious diseases and zoonoses. Zoonoses are defined as infectious diseases that are spread between animals and humans through different mechanisms, including direct contact, food, water, vectors, or fomites. This review highlights examples in which human and veterinary pathologists were an integral part of the multisectoral team that identified uncommon etiologic agents or pathologies that had not been elucidated clinically. As the team discovers an emerging infectious disease, pathologists develop and validate tests for epidemiologic and clinical use and provide surveillance data on these diseases. They define the pathogenesis and pathology that these new diseases cause. This review also presents examples that demonstrate the crucial role pathologists play in diagnosing zoonoses that have an impact on the food supply and the economy.

We performed an epidemiological investigation and genome sequencing of severe acute respiratory coronavirus virus 2 (SARS-CoV-2) to define the source and scope of an outbreak in a cluster of hospitalized patients. Lack of appropriate respiratory hygiene led to SARS-CoV-2 transmission to patients and healthcare workers during a single hemodialysis session, highlighting the importance of infection prevention precautions.

Abstract Monitoring drug response to ensure optimal platelet suppression is necessary in select patients on dual antiplatelet therapy (DAPT). At our institution, responses to aspirin (ASA) and P2Y12 inhibitors (P2Y12-Is) are assessed by light transmission aggregometry (LTA) using the Helena AggRAM assay. This method requires platelet-rich plasma (PRP) and is labor intensive. Platelet mapping using thromboelastography (TEG) is an alternate method, which offers point-of-care testing on whole blood. Here, we aimed to evaluate the concordance of our LTA assay with TEG, as well as the VerifyNow® PRUTest. We aimed to run testing on at least 20 patient samples; however, at the time of abstract submission, only 10 samples were available. Ten blood samples from 8 patients on dual anti-platelet therapy were collected in preparation for neurologic stent placement. Prepared PRP was analyzed by LTA on the Helena AggRAM assay. The maximal amplitude of the aggregation curves was recorded in the presence of high dose (HD) ADP, low dose (LD) ADP, and arachidonic acid (AA). Whole blood was analyzed by TEG using PlateletMapping ADP & AA assay cartridges on the Haemonetics® TEG 6s analyzer, and by the VerifyNow® PRUTest. Thresholds to qualify platelet suppression as optimal or suboptimal were defined a priori following established guidelines. Concordance between methods was calculated using the following formula: (optimal/optimal + suboptimal/suboptimal) / total number of samples. Pearson correlation between units of measure was also determined. For P2Y12-I response, concordance between LTA and TEG was 70% (correlation was r = 0.76 between TEG and LTA with HD ADP, and r = 0.70 between TEG and LTA with LD ADP). Concordance between VerifyNow® and LTA was 60% (correlation was r = 0.84 between VerifyNow® and LTA with HD ADP, and r = 0.78 between VerifyNow® and LTA with LD ADP). Finally, concordance between VerifyNow® and TEG was 30% (correlation was r = 0.81). Regarding ASA response, concordance was 80% between LTA and TEG (correlation was r = 0.31). To summarize, overall concordance ranged from 30 to 80% across platforms. For P2Y12 inhibition, results obtained from LTA with HD ADP and VerifyNow® had the strongest correlation (r = 0.84). Interestingly, the correlation between VerifyNow® and TEG was similarly elevated (r = 0.81), but concordance was poor (30%). When comparing TEG to LTA, the strongest correlation was between LTA with HD ADP and TEG (r value = 0.76). Concerning ASA inhibition, concordance between LTA and TEG was acceptable; however, correlation was weak (80%; r = 0.31). Results from this preliminary study support the potential use of TEG to monitor platelet response to therapy. We plan to continue our testing to include a minimum of 20 samples, and also consider patient outcomes, before implementing any laboratory workflow changes.

OBJECTIVES:Provide quality management training in anatomic pathology so that slides are of adequate quality and can be interpreted.METHODS:During the first African Pathology Assembly, we performed a needs assessment and knowledge quizzes, then presented 4 modules of the quality management system (personnel management, process control, sample management, and equipment) that are used to train quality in vertical programs by the World Health Organization.RESULTS:Participants included 14 (34%) trainees, 14 (34%) pathologists, and 9 (22%) technologists from South Africa (11), Nigeria (6), Tanzania (4), and other countries (18). Thirty (73%) participants took the course because they had interest in the topic while 6 (15%) did it because it was recommended by a supervisor. Most participants thought that the quality of slides was medium to high in their institution and that clinicians trust results. The most frequent quality issues cited included problems from processing to staining, long turnaround times, and preanalytical issues (fixation, lack of clinical history). The average result of the knowledge quiz was 6.7 (range, 2-10) before (38 participants) the course and 8.3 (range, 5-10) after (30 participants) the course.CONCLUSIONS:This assessment suggests there is a need for quality management courses in pathology in Africa.

The burden of emerging antimicrobial resistance (AMR) in the United States is significant and even greater worldwide. Mitigation efforts have decreased the incidence and deaths from antimicrobial-resistant organisms in the United States. Yet more than 2.8 million antimicrobial-resistant infections occur every year and more than 35,000 patients die as a result. Infection prevention and control, data tracking, antimicrobial stewardship, vaccines, therapeutics, diagnostics, and sanitation are all required to decrease AMR threats. In 2019, in the second version of the Centers for Disease Control and Prevention (CDC) report on antibiotic-resistant threats, the agency categorized AMR threats as urgent, serious, concerning, or to be watched. This review will discuss the following aspects of each bacterium in the CDC report: estimated numbers of cases and deaths, identify the better known and impactful mechanisms of resistance, diagnostic testing and its limitations, and current and possible future therapies. This review also presents anatomical pathology case examples that highlight the altered morphology of antibiotic partially treated bacteria in tissues.

Traditional cellular and live-virus methods for detection of SARS-CoV-2 neutralizing antibodies (nAbs) are labor- and time-intensive, and thus not suited for routine use in the clinical lab to predict vaccine efficacy and natural immune protection. Here, we report the development and validation of a rapid, high throughput method for measuring SARS-CoV-2 nAbs against native-like trimeric spike proteins. This assay uses a blockade of human angiotensin converting enzyme 2 (hACE-2) binding (BoAb) approach in an automated digital immunoassay on the Quanterix HD-X platform. BoAb assays using Wuhan-WT (vaccine strain), delta (B.1.167.2), omicron BA1 and BA2 variant viral strains showed strong correlation with cell-based pseudovirus neutralization activity (PNA) and live-virus neutralization activity. Importantly, we were able to detect similar patterns of delta and omicron variant resistance to neutralization in samples with paired vaccine strain and delta variant BoAb measurements. Finally, we screened clinical samples from patients with or without evidence of SARS-CoV-2 exposure by a single-dilution screening version of our assays, finding significant nAb activity only in exposed individuals. Importantly, this completely automated assay can be performed in 4 h to measure neutralizing antibody titers for 16 samples over 8 serial dilutions or, 128 samples at a single dilution with replicates. In principle, these assays offer a rapid, robust, and scalable alternative to time-, skill-, and cost-intensive standard methods for measuring SARS-CoV-2 nAb levels.