CE-MS-Based Clinical Metabolomics of Human Plasma.

Capillary electrophoresis coupled to mass spectrometry (CE-MS) has emerged as a powerful analytical technique with significant implications for clinical research and diagnostics. The integration of information from CE and MS strengthens confidence in the identification of compounds present in clinical samples. The ability of CE to separate molecules based on their electrophoretic mobility coupled to MS enables the accurate identification and quantification of analytes, even in complex biological matrices such as human plasma.Here, we present a detailed protocol for an untargeted metabolomics study using CE-MS and its application in a study on human plasma from patients suffering Long COVID syndrome. The protocol ranges from sample preparation to biological interpretation, detailing a workflow enabling the analysis of cationic and anionic compounds, metabolite identification, and data processing.

Accuracy study of Angiotensin 1-7 composite index test to predict pulmonary fibrosis and guide treatment.

BACKGROUND: Pulmonary fibrosis can develop after acute respiratory distress syndrome (ARDS). The hypothesis is we are able to measure phenotypes that lie at the origin of ARDS severity and fibrosis development. The aim is an accuracy study of prognostic circulating biomarkers. METHODS: A longitudinal study followed COVID-related ARDS patients with medical imaging, pulmonary function tests and biomarker analysis, generating 444 laboratory data. Comparison to controls used non-parametrical statistics; p < 0·05 was considered significant. Cut-offs were obtained through receiver operating curve. Contingency tables revealed predictive values. Odds ratio was calculated through logistic regression. RESULTS: Angiotensin 1-7 beneath 138 pg/mL defined Angiotensin imbalance phenotype. Hyper-inflammatory phenotype showed a composite index test above 34, based on high Angiotensin 1-7, C-Reactive Protein, Ferritin and Transforming Growth Factor-ß. Analytical study showed conformity to predefined goals. Clinical performance gave a positive predictive value of 95 % (95 % confidence interval, 82 %-99 %), and a negative predictive value of 100 % (95 % confidence interval, 65 %-100 %). Those severe ARDS phenotypes represented 34 (Odds 95 % confidence interval, 3-355) times higher risk for pulmonary fibrosis development (p < 0·001). CONCLUSIONS: Angiotensin 1-7 composite index is an early and objective predictor of ARDS evolving to pulmonary fibrosis. It may guide therapeutic decisions in targeted phenotypes.

Thiamine-responsive megaloblastic anaemia in a young adult with acute pancytopenia.

Thiamine-responsive megaloblastic anaemia (TRMA) is a rare autosomal recessive disorder characterised by the clinical triad of megaloblastic anaemia, sensorineural hearing loss and diabetes mellitus (DM) in young patients. We present a case of a young man with type 1 DM who presented with pancytopenia of unclear aetiology, initially attributed to a COVID-19 infection. After obtaining a bone marrow biopsy and pursuing genetic testing, two pathogenic variants of the SLC19A2 gene consistent with TRMA were discovered in this patient. Treatment with 100 mg of thiamine oral supplementation daily led to the complete resolution of his pancytopenia. It is important to consider a genetic cause of pancytopenia in a young person. Early recognition and diagnosis of TRMA can be life-altering given early treatment can reduce insulin requirements and resolve anaemia.

Four Models of Wastewater-Based Monitoring for SARS-CoV-2 Complementing Individual Screening in Jail Settings.

Objectives. To describe 4 unique models of operationalizing wastewater-based surveillance (WBS) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in jails of graduated sizes and different architectural designs. Methods. We summarize how jails of Cook County, Illinois (average daily population [ADP] 6000); Fulton County, Georgia (ADP 3000); Middlesex County, Massachusetts (ADP 875); and Washington, DC (ADP 1600) initiated WBS between 2020 and 2023. Results. Positive signals for SARS-CoV-2 via WBS can herald a new onset of infections in previously uninfected jail housing units. Challenges implementing WBS included political will and realized value, funding, understanding the building architecture, and the need for details in the findings. Conclusions. WBS has been effective for detecting outbreaks of SARS-CoV-2 in different sized jails, those with both dorm- and cell-based architectural design. Public Health Implications. Given its effectiveness in monitoring SARS-CoV-2, WBS provides a model for population-based surveillance in carceral facilities for future infectious disease outbreaks. (Am J Public Health. 2024;114(11):1232-1241. https://doi.org/10.2105/AJPH.2024.307785).

Branch retinal artery occlusion in a patient with COVID-19 infection: A case report.

We report here, a case of branch retinal artery occlusion (BRAO) in the left eye of a 76-year-old man that involved three arteries which was considered to be related to a COVID-19 infection due to high levels of blood cytokines and coagulation factors. Although the patient had hypertension and atherosclerosis, his hypertension had been well controlled for the past five years by regular antihypertensive medication. Twenty-five days after starting treatment with anti-inflammatory, anticoagulant and conservative therapy, the patient's biomarkers of inflammation and coagulation returned to normal and his vision improved. However, some visual field defects remained and were probably a consequence of low oxygen saturation.

Impact of SARS-CoV-2 viral load on restrictive spirometry patterns in mild COVID-19 recovered middle-aged individuals: a six-month prospective study.

BACKGROUND: Long term respiratory complications of Corona Virus Disease-2019 (COVID-19) are of great concern. Many studies have reported altered respiratory patterns in COVID-19 recovered individuals and most of them were from severe to critically ill patients. The association of viral load at the time of infection with symptoms of long COVID-19 specifically on pulmonary functions after months of recovery is still not known. This study was aimed to assess the impact of SARS-CoV-2 viral load during mild-moderate COVID-19 disease on pulmonary functions in middle-aged population after 6-8 months of acute infection. METHODS: This study included 300 (102 healthy controls and 198 COVID-19 recovered) individuals between age 30-60 of either gender. Mild-moderate COVID-19 recovered individuals were recruited between a period of 6-8 months post-acute infection. Spirometry was performed with MIR-Spirolab-III. The association of spirometry pattern was compared with SARS-CoV-2 viral loads during acute infection. RESULTS: We observed up to 70% of the participants presented with either shortness of breath (11.5%), body aches (23.5%), recurrent cough (4.4%), recurrent respiratory infections (9.5%) and/or fatigue (33.3%) at follow up. In our study, 35.5% of COVID-19 recovered individuals had abnormal respiratory patterns (33.5% had restrictive and 2% had obstructive patterns). Viral load ≤ 20 CT value was associated with restrictive respiratory patterns (p = 0.004). No association was found between viral load and disease severity (p = 0.23). CONCLUSION: In this study, we found one third of mild-moderate COVID-19 recovered individuals have restrictive respiratory patterns after 6-8 months of recovery. These findings had a strong association with SARS-CoV-2 viral loads during acute infection which has been reported for the first time in our study. Studying the relationship between viral load and pulmonary functions can contribute to identifying potential risk factors for long COVID and developing preventive measures to mitigate the long-term impact on lung health. CLINICAL TRIAL NUMBER: Not applicable.

Performance of the 4C and SEIMC scoring systems in predicting mortality from onset to current COVID-19 pandemic in emergency departments.

The evolution of the COVID-19 pandemic has been associated with variations in clinical presentation and severity. Similarly, prediction scores may suffer changes in their diagnostic accuracy. The aim of this study was to test the 30-day mortality predictive validity of the 4C and SEIMC scores during the sixth wave of the pandemic and to compare them with those of validation studies. This was a longitudinal retrospective observational study. COVID-19 patients who were admitted to the Emergency Department of a Spanish hospital from December 15, 2021, to January 31, 2022, were selected. A side-by-side comparison with the pivotal validation studies was subsequently performed. The main measures were 30-day mortality and the 4C and SEIMC scores. A total of 27,614 patients were considered in the study, including 22,361 from the 4C, 4,627 from the SEIMC and 626 from our hospital. The 30-day mortality rate was significantly lower than that reported in the validation studies. The AUCs were 0.931 (95% CI: 0.90-0.95) for 4C and 0.903 (95% CI: 086-0.93) for SEIMC, which were significantly greater than those obtained in the first wave. Despite the changes that have occurred during the coronavirus disease 2019 (COVID-19) pandemic, with a reduction in lethality, scorecard systems are currently still useful tools for detecting patients with poor disease risk, with better prognostic capacity.

Lateral flow assay sensitivity and signal enhancement via laser µ-machined constrains in nitrocellulose membrane.

Lateral flow assay (LFA) is a handful diagnostic technology that can identify severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other common respiratory viruses in one strip, which can be tested at the point-of-care without the need for equipment or skilled personnel outside the laboratory. Although its simplicity and practicality make it an appealing solution, it remains a grand challenge to substantially enhance the colorimetric LFA sensitivity. In this work, we present a straightforward approach to enhance the sensitivity of LFA by imposing the flow constraints in nitrocellulose (NC) membranes via a number of vertical femtosecond laser micromachined microchannels which is important for prolonged specific binding interactions. Porous NC membrane surfaces were structured with different widths and densities µ-channels employing a second harmonic of the Yb:KGW femtosecond laser and sample XYZ translation over a microscope objective-focused laser beam. The influence of the microchannel parameters on the vertical wicking speed was evaluated from the video recordings. The obtained results indicated that µ-channel length, width, and density in NC membranes controllably increased the immunological reaction time between the analyte and the labeled antibody by 950%. Image analysis of the colorimetric indicators confirmed that the flow rate delaying strategy enhanced the signal sensitives by 40% compared with pristine NC LFA.

Diagnostics for detection and surveillance of priority epidemic-prone diseases in Africa: an assessment of testing capacity and laboratory strengthening needs.

In 2023, Africa experienced 180 public health emergencies, of which 90% were infectious diseases and 75% were related to zoonotic diseases. Testing capacity for epidemic-prone diseases is essential to enable rapid and accurate identification of causative agents, and for action to prevent disease spread. Moreover, testing is pivotal in monitoring disease transmission, evaluating public health interventions and informing targeted resource allocation during outbreaks. An online, self-assessment survey was conducted in African Union Member States to identify major challenges in testing for epidemic-prone diseases. The survey assessed current capacity for diagnosing priority epidemic-prone diseases at different laboratory levels. It explored challenges in establishing and maintaining testing capacity to improve outbreak response and mitigate public health impact. Survey data analysed diagnostic capacity for priority infectious diseases, diagnostic technologies in use, existing surveillance programmes and challenges limiting diagnostic capacity, by country. The survey result from 15 Member States who responded to the survey, showed high variability in testing capacity and technologies across countries and diverse factors limiting testing capacity for certain priority diseases like dengue and Crimean-Congo haemorrhagic fever. At the same time diagnostic capacity is better for coronavirus disease 2019 (COVID-19), polio, and measles due to previous investments. Unfortunately, many countries are not utilizing multiplex testing, despite its potential to improve diagnostic access. The challenges of limited laboratory capacity for testing future outbreaks are indeed significant. Recent disease outbreaks in Africa have underscored the urgent need to strengthen diagnostic capacity and introduce cost-effective technologies. Small sample sizes and differing disease prioritisation within each country limited the analysis. These findings suggest the benefits of evaluating laboratory testing capacity for epidemic-prone diseases and highlight the importance of effectively addressing challenges to detect diseases and prevent future pandemics.

LAMP-CRISPR/Cas12a-based impedimetric biosensor powered by Fe3O4@Au-(S-polyA-S)-Au for detection of SARS-CoV-2.

A low-cost, lab-made polytetrafluoroethylene micro-cell, equipped with three electrodes, wasd eveloped for the impedimetric detection of SARS-CoV-2. The gold working electrode was modified with a double-ended thiolated poly-adenine probe, which was conjugated with magnetic Fe3O4@Au nanoparticles (Fe3O4@Au-(S-polyA-S)-Au). After the loop-mediated isothermal amplification (LAMP) of viral RNA, the single-guide RNA (sgRNA), specifically bound to the SARS-CoV-2 target sequence, activates Cas12a. Cas12a then cleaved the immobilized probe. As a result, the magnetic Fe3O4@Au nanoparticles were released and adsorbed onto the gold electrode surface, using an external magnet. This process increased the physical surface area of the gold electrode, facilitating redox ion ([FeIII/II(CN)6]3-/4-) electron transfer. The decrease in the charge transfer resistance was utilized for SARS-CoV-2 detection. Our LAMP-CRISPR/Cas12a-based impedimetric biosensor, powered by Fe3O4@Au-(S-polyA-S)-Au, demonstrated impressive capabilities, including a remarkable detection limit of 0.8 aM (0.48 copies/µL) and a linear range of 0.01 to 36.06 fM.

Using digital tools and antigen rapid testing to support household-level SARS-CoV-2 detection by community health workers in Rwanda: an operational pilot study.

OBJECTIVE: To evaluate the use of antigen-based rapid diagnostic tests (Ag-RDTs) alongside a digital tool to deliver household-level COVID-19 testing by community health workers (CHWs), in line with Rwanda's ambition to decentralise COVID-19 testing. DESIGN: This was an operational pilot study to evaluate the impact and operational characteristics of using the digital e-ASCov tool combined with Ag-RDTs to support COVID-19 symptom screening and rapid testing by CHWs across eight districts in Rwanda. A total of 800 CHWs selected from both rural and urban areas were trained in delivering Ag-RDTs for COVID-19 testing and using the e-ASCOV application for data capture on a smartphone. Laboratory technicians repeated a subset of Ag-RDTs to assess the concordance of results obtained by CHWs. The study also assessed CHWs' experience of the intervention using a mixed-methods approach. SETTING: Eight rural, urban and semiurban districts in Rwanda. PARTICIPANTS: A total of 19 544 individuals were enrolled and screened for signs and symptoms of COVID-19. INTERVENTIONS: Community-based screening for COVID-19 by CHWs using the digital tool e-ASCov combined with rapid testing using Ag-RDTs. MAIN OUTCOME MEASURES: Number of participants screened and tested; concordance of Ag-RDT results between CHWs and laboratory technicians; feasibility of study procedures by CHWs and CHWs perceptions of the digital tool and Ag-RDT testing. RESULTS: From February to May 2022, CHWs screened 19 544 participants, of whom 4575 (23.4%) had COVID-19-related symptoms or a history of exposure to the infection. Among them, 86 (1.9%) were positive on Ag-RDTs. Concordance of Ag-RDT results between CHWs and laboratory technicians was 100%. Of the 800 trained CHWs, 746 (93.3%) were independently able to conduct household-based COVID-19 screening, perform the Ag-RDTs and send data to the central server. Most CHWs (>80%) found Ag-RDTs and e-ASCOV easy to use. CONCLUSIONS: This study demonstrated the feasibility of deploying a digital tool and Ag-RDTs for household-level SARS-CoV-2 detection in Rwanda. The findings support a broader roll-out of digitally supported rapid testing by CHWs to broaden access to testing for priority diseases.

Assessing the impact of COVID-19 on tuberculosis detection and treatment in healthcare facilities across Addis Ababa, Ethiopia: A comprehensive mixed-method, multi-center study.

BACKGROUND: Ethiopia faces a significant burden of Tuberculosis (TB), being one of the high-burden countries, and the emergence of the Coronavirus Disease 2019 (COVID-19) has become a dominant health concern, particularly in resource-limited settings. The repercussions of COVID-19 on TB care are evident, leading to a surge in undiagnosed TB cases, challenges in medication adherence, and an escalation of drug resistance. Consequently, a thorough assessment of the impact of COVID-19 on TB care becomes imperative to devise a tailored program for managing TB amidst future pandemics, natural disasters, and conflict crises. METHODS: A mixed-methods study design was utilized, encompassing a randomly selected 10 health centers (HCs) and 3 hospitals among government owned 98 HCs and 5 hospitals in Addis Ababa, Ethiopia. All TB patients who were on follow-up during the study period were included. The study period was from March 4, 2020, to December 4, 2020, with the corresponding period of March 4, 2019, to December 4, 2019, serving as the baseline for comparison. Quantitative data were gathered from TB patients' medical registries, laboratory registries, and treatment follow-up charts. Complementary qualitative data were acquired through in-depth interviews. Both qualitative and quantitative data were collected from January 17, 2022 to May 13, 2022. RESULTS: Following the onset of the pandemic, there was a notable and statistically significant decline in both the detection of TB cases and the number of positive results across all study sites. Bacteriological TB tests reduced from 5837 to 2126 patients, and TB-positive cases decreased from 500 to 218, representing declines of 63.6% and 56.4%, respectively. The overall number of TB patients undergoing treatment also experienced a decrease from 1431 to 1051, marking a 26.6% reduction. Additionally, there was a 10% increase in the proportion of extra-pulmonary TB cases. The impact of the pandemic extended to TB treatment outcomes, with adverse effects on cure rates, death rates, loss of follow-up, and medication adherence. The apprehension of contracting COVID-19 and the implementation of isolation measures contributed to a decline in healthcare-seeking behaviors among patients, fostering negative perceptions and practices among healthcare workers. The challenges further exacerbated due to a shortage of personal protective equipment, a lack of rapid diagnostic test tools, clinical presentations resembling COVID-19, and a shift in government policies. These factors collectively posed significant obstacles to effective TB care during the pandemic. CONCLUSION: The profound impact of COVID-19 on critical TB care service indicators, including TB detection, treatment initiation, and treatment outcomes, underscores the need for immediate and collaborative measures. It is imperative to implement strategies that ensure the resumption of all TB care services concurrently with efforts to control COVID-19. A comprehensive and coordinated approach is essential to mitigate the adverse effects of the pandemic on TB management and safeguard public health.

Oral SARS-CoV-2 host responses predict the early COVID-19 disease course.

Oral fluids provide ready detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and host responses. This study sought to evaluate relationships between oral virus, oral and systemic anti-SARS-CoV-2-specific antibodies, and symptoms. Oral fluids (saliva/throat wash (saliva/TW)) and serum were collected from asymptomatic and symptomatic, nasopharyngeal (NP) SARS-CoV-2 RT-qPCR+ human participants (n = 45). SARS-CoV-2 RT-qPCR and N-antigen detection by immunoblot and lateral flow assay (LFA) were performed. RT-qPCR for subgenomic RNA (sgRNA) was sequence confirmed. SARS-CoV-2-anti-S protein RBD LFA and ELISA assessed IgM and IgG responses. Structural analysis identified host salivary molecules analogous to SARS-CoV-2-N-antigen. At time of enrollment (baseline, BL), LFA-detected N-antigen in 86% of TW and was immunoblot-confirmed. However, only 3/17 were saliva/TW qPCR+ . Sixty percent of saliva and 83% of TW demonstrated persistent N-antigen at 4 weeks. N-antigen LFA signal in three anti-spike sero-negative participants suggested potential cross-detection of 4 structurally analogous salivary RNA binding proteins (alignment 19-29aa, RMSD 1-1.5 Angstroms). At enrollment, symptomatic participants demonstrated replication-associated sgRNA junctions, were IgG+ (94%/100% in saliva/TW), and IgM+ (63%/54%). At 4 weeks, SARS-CoV-2 IgG (100%/83%) and IgM (80%/67%) persisted. Oral and serum IgG correlated 100% with NP+ PCR status. Cough and fatigue severity (p = 0.010 and 0.018 respectively), and presence of weakness, nausea, and composite upper respiratory symptoms (p = 0.037, 0.005, and 0.017, respectively) were negatively associated with saliva IgM but not TW or serum IgM. Throat wash IgM levels were higher in women compared to men, although the association did not reach statistical significance (median: 290 (female) versus 0.697, p = 0.056). Important to transmission and disease course, oral viral replication and persistence showed clear relationships with select symptoms and early oral IgM responses during early infection. N-antigen cross-reactivity may reflect mimicry of structurally analogous host proteins.

Acceptance of SARS-CoV-2 Surveillance Testing Among Patients Receiving Dialysis: A Cluster Randomized Trial.

Importance: Integrating routine SARS-CoV-2 testing in dialysis facilities may benefit patients receiving dialysis by mitigating risks of serious illness and reducing transmission. Patient acceptance of nonmandatory testing is unknown. Objective: To evaluate the acceptance of 2 SARS-CoV-2 testing strategies among patients in hemodialysis facilities nationwide. Design, Setting, and Participants: This nationwide cluster (dialysis facility-level) randomized trial investigated the acceptance of SARS-CoV-2 testing among patients receiving maintenance hemodialysis at facilities located in 22 states. Intervention: Anterior nares real-time reverse transcriptase-polymerase chain reaction tests offered once every 2 weeks (static testing facilities) vs offered once a week, once every 2 weeks, or once a month depending on county COVID-19 infection prevalence (dynamic testing facilities). Facilities were randomized by county, and tests were offered for 3 months between February 4 and July 24, 2023. Main Outcomes and Measures: The primary outcome was test acceptance. Secondary outcomes included the proportion of patients who accepted at least 1 test. Results: In total, 62 hemodialysis facilities were randomized and 57 participated. Among 2389 participating patients, the median age was 64 (IQR, 54-74) years, 1341 (56%) were male, 138 (6%) were categorized as American Indian, 60 (3%) Asian, 885 (37%) Black, 75 (3%) Native Hawaiian or Pacific Islander, 338 (14%) Hispanic, and 876 (37%) White; and 1603 (67%) had diabetes. A median of 6 (IQR, 6-6) tests were offered per patient in the static arm and 4 (3-6) tests in the dynamic arm. Test acceptance was low: 8% of offered tests were accepted in each of the test arms. Among 503 patients who accepted at least 1 test, the median percentage of offered tests that were accepted was 16% (IQR, 17%-42%) using the static testing strategy and 50% (IQR, 33%-75%) using the dynamic testing strategy (P < .001). Older patients (odds ratio [OR], 1.08 [95% CI, 1.01-1.16] per 5-year age increment), patients with (vs without) diabetes (OR, 1.59 [95% CI, 1.18-2.16]), and women compared with men (OR, 1.30 [95% CI, 0.98-1.73]) were more likely to accept multiple tests. Patients designated in the electronic health record as Hispanic were more likely than patients designated as White (OR, 1.78 [95% CI, 1.15-2.76]) to accept at least 1 test, whereas patients living in zip codes electing Republican representatives to Congress were less likely than patients living in zip codes electing Democratic representatives (OR, 0.34 [95% CI, 0.17-0.69]) to accept multiple tests. Conclusions and Relevance: In this cluster randomized trial evaluating 2 SARS-CoV-2 testing strategies in dialysis facilities, test acceptance was low, and a dynamic testing strategy anchored to COVID-19 infection prevalence did not outperform a static testing strategy of every 2 weeks. Trial Registration: ClinicalTrials.gov Identifier: NCT05225298.

Utilising accessible and reproducible neurological assessments in clinical studies: Insights from use of the Neurological Impairment Scale in the multi-centre COVID-CNS study.

Reproducible and standardised neurological assessment scales are important in quantifying research outcomes. These scales are often performed by non-neurologists and/or non-clinicians and must be robust, quantifiable, reproducible and comparable to a neurologist's assessment. COVID-CNS is a multi-centre study which utilised the Neurological Impairment Scale (NIS) as a core assessment tool in studying neurological outcomes following COVID-19 infection. We investigated the strengths and weaknesses of the NIS when used by non-neurology clinicians and non-clinicians, and compared performance to a structured neurological examination performed by a neurology clinician. Through our findings, we provide practical advice on how non-clinicians can be readily trained in conducting reproducible and standardised neurological assessments in a multi-centre study, as well as illustrating potential pitfalls of these tools.

COVID-19 diagnosis on the basis of nanobiosensors' prompt interactivity: A holistic review.

The fast spread and severe consequences of novel coronavirus disease 2019 (COVID-19) have once again underscored the critical necessity of early detection of viral infections. Several serology-based techniques, including as point-of-care assays and high-throughput enzyme immunoassays that support the diagnosis of COVID-19 are utilized in the detection and identification of coronaviruses. A rapid, precise, simple, affordable, and adaptable diagnostic tool is required for controlling COVID-19 as well as for outbreak management, since the calculation and monitoring of viral loads are crucial for predicting the infection stage and recovery time. Nowadays, the most popular method for diagnosing COVID-19 is reverse transcription polymerase chain reaction (RT-PCR) testing, and chest computed tomography (CT) scans are also used to determine the disease's phases. This is all because of the fact that RT-PCR method caries with itself a number of downsides comprising of being immovable, expensive, and laborious. RT-PCR has not well proven to be capable of detection on the very early infection stages. Nanomaterial-based diagnostics, together with traditional clinical procedures, have a lot of promise against COVID-19. It is worthy of attention that nanotechnology has the mainstay capacity for purposes of developing even more modern stratagems fighting COVID-19 by means of focusing on state-of-the-art diagnostics. What we have centered on in this review, is bringing out even more efficient detection techniques whereby nanobiosensors are employed so that we might obstruct any further development and spreading of SARS-CoV-2.

Disparities in age and gender-specific SARS-CoV-2 diagnostic testing trends: a retrospective study from Pakistan.

BACKGROUND: Pakistan reported 1.57 million COVID-19 cases between 2020 and 2022, based on approximately 30.6 million SARS-CoV-2 RT-PCR (reverse-transcription polymerase chain reaction) tests conducted. This study utilized data from one of the largest in-country testing facilities, Aga Khan University Hospital (AKUH) in Karachi, Pakistan, to explore gender and age-related in RT-PCR testing patterns. METHODS: We conducted a retrospective review of SARS-CoV-2 RT-PCR test data extracted from AKUH clinical laboratory records between February 2020 and February 2022. Gender and age distributions were examined in the context of testing patterns across the period. Multivariate regression models assessed independent associations between COVID-19 positivity and key variables. RESULTS: We reviewed 470,249 RT-PCR tests, finding that most tests were in those aged 21-40 years (48.1%). Overall, COVID-19 test positivity was 20.6%. In all, 57.7% were performed for males, predominant amongst those tested across all age groups and waves. Females had significantly lower odds of testing positive for COVID-19 (OR: 0.9; 95% CI: 0.9-1.0). However, when adjusted for gender, age and pandemic phases, the positivity rates between males and females were the same. The odds of a positive result increased significantly with age; individuals aged > 80 years had 2.5 times higher odds of testing positive than those aged 0-10 years (aOR 2.5, 95% CI 2.3-2.7). CONCLUSIONS: The analysis indicates a consistent male dominance in COVID-19 testing, with higher positivity rates in older age groups. Our study highlight the importance of examining demographic characteristics in disease associated data especially, representation of females amongst cohorts.

Predicting malignant potential of solitary pulmonary nodules in patients with COVID-19 infection: a comprehensive analysis of CT imaging and tumor markers.

OBJECTIVE: To analyze the value of combining computed tomography (CT) with serum tumor markers in the differential diagnosis of benign and malignant solitary pulmonary nodules (SPNs). METHODS: The case data of 267 patients diagnosed with SPNs in the First Affiliated Hospital of Zhengzhou University from March 2020 to January 2023 were retrospectively analyzed. All individuals diagnosed with coronavirus disease 2019 (COVID-19) were confirmed via respiratory specimen viral nucleic acid testing. The included cases underwent CT, serum tumor marker testing and pathological examination. The diagnostic efficacy and clinical significance of CT, serum tumor marker testing and a combined test in identifying benign and malignant SPNs were analyzed using pathological histological findings as the gold standard. Finally, a nomogram mathematical model was established to predict the malignant probability of SPNs. RESULTS: Of the 267 patients with SPNs, 91 patients were not afflicted with COVID-19, 36 exhibited malignant characteristics, whereas 55 demonstrated benign features. Conversely, within the cohort of 176 COVID-19 patients presenting with SPNs, 62 were identified as having malignant SPNs, and the remaining 114 were diagnosed with benign SPNs. CT scans revealed statistically significant differences between the benign and malignant SPNs groups in terms of CT values (P<0.001), maximum nodule diameter (P<0.001), vascular convergence sign (P<0.001), vacuole sign (P = 0.0007), air bronchogram sign (P = 0.0005), and lobulation sign (P = 0.0005). Malignant SPNs were associated with significantly higher levels of carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) compared to benign SPNs (P < 0.05), while no significant difference was found in carbohydrate antigen 125 (CA125) levels (P = 0.054 for non-COVID-19; P = 0.072 for COVID-19). The sensitivity (95.83%), specificity (95.32%), and accuracy (95.51%) of the comprehensive diagnosis combining serum tumor markers and CT were significantly higher than those of CT alone (70.45%, 79.89%, 76.78%) or serum tumor marker testing alone (56.52%, 73.71%, 67.79%) (P < 0.05). A visual nomogram predictive model for malignant pulmonary nodules was constructed. CONCLUSION: Combining CT with testing for CEA, CA125, and NSE levels offers high diagnostic accuracy and sensitivity, enables precise differentiation between benign and malignant nodules, particularly in the context of COVID-19, thereby reducing the risk of unnecessary surgical interventions.

Clinical characteristics and outcomes of neonatal SARS-CoV-2 infection after the release of the epidemic situation of COVID-19.

BACKGROUND: With the release of the coronavirus disease 2019 (COVID-19) pandemic in late 2022 in China, the number of people infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) roared, including neonatal cases. However, there were few studies on neonatal COVID-19, especially multi-center case reports. This study aimed to explore clinical characteristics and short-term outcomes of neonatal COVID-19 in China. METHODS: We reviewed 187 cases of neonatal COVID-19 between December 11, 2022, and January 12, 2023. The diagnosis was assessed by symptoms, laboratory tests, X-ray manifestations, and diagnosis code. Clinical characteristics and outcomes were evaluated. RESULTS: In 187 neonatal cases with COVID-19, 84 (44.9%) had severe SARS-CoV-2 infection. Most patients had confirmed exposure to SARS-CoV-2. Fever and respiratory symptoms were common (75.4% and 71.7%, respectively). Severe patients were more likely to have high alanine transaminase (ALT) (> 40U/L) (11.9% vs. 3.9%) and high N-terminal pro-brain natriuretic peptide (NT-proBNP) (> 2000pg/mL) (38.0% vs. 19.6%), compared with nonsevere ones (P < 0.05). None of the patients received COVID-19-specific medical interventions. A few severe patients received corticosteroids (1.1%), and immunoglobulin (0.5%), respectively. All patients were discharged home after the medical care with a median length of stay (LOS) of four days and none of them met the criteria of multisystem inflammatory syndrome in neonates (MIS-N). CONCLUSIONS: After the release of the epidemic situation of COVID-19 in late 2022 in China, more neonatal cases with severe COVID-19 had high ALT and NT-proBNP level. Few specific medical interventions were given, and the outcome was satisfying.