Journal of Medical Virology最新文献

The global impact of pandemics has intensified the demand for rapid, cost-effective, sensitive, and reliable POCT methods. Herein, we developed a multiplex pathogen detection system based on a dual-drive microfluidic chip, enabling simultaneous detection of RSV, FluA, FluB, and SARS-CoV-2 antigens from a single sample via parallel-arranged detection units within microchannels. Monoclonal antibodies targeting conserved viral proteins were cross-paired to identify optimal pairs. The chip's labeling and reference regions were designed, and key parameters were optimized to enhance performance. Cut-off values were established using nasopharyngeal swab samples from healthy donors, and assay performance was rigorously validated. Finally, the concordance between the multiplex chip and GICA and RT-PCR was evaluated. The optimized process ultimately yielded a chip design featuring a labeling region with four antiviral monoclonal antibody-conjugated fluorescent microspheres, four test regions each coated with corresponding antiviral monoclonal antibodies, and a dual-antibody reference region for signal normalization. The developed assay requires only 35 μL of sample and completes detection within 5 min. It demonstrated high repeatability, with coefficients of variation ranging from 9.48% to 13.37%. The detection limits were 0.625 ng/mL for RSV, 2.5 ng/mL for FluA, 5 ng/mL for both FluB and SARS-CoV-2. Validation studies showed perfect agreement with GICA (n = 132, Kappa = 1) and strong consistency with RT-PCR (Kappa = 0.74–0.90, n = 42). A novel POCT method has been successfully established for the simultaneous detection of four respiratory virus antigens. This system shows great potential as a reliable and efficient analytical tool for large-scale epidemic screening.

Enteroviruses (EVs) cause a spectrum of illnesses ranging from mild to severe, including neurological complications. In this study, the molecular epidemiology, clinical impact, and seasonality of EVs were evaluated among hospitalized patients in a large tertiary medical center in Israel from 2016 to 2024. A total of 11,246 cerebrospinal fluid (CSF) samples and 5,744 stool samples were tested for EV RNA using RT-PCR, with 7.1% (n = 798) and 19.3% (n = 1,110) testing positive, respectively. Positive samples were sequenced for genotype identification based on partial VP1 sequences with 70% genotype identification. The most commonly detected subtypes in CSF samples were: E-18 (16.4%), E-5 (80/567, 14.1%), E-6 (67/567, 11.8%), E-30 (64/567, 11.3%), CVB5 (37/567, 6.5%), E-4 (29/567, 5.1%), and CVB2 (28/567, 4.9%). These subtypes were also detected in stool samples during similar time periods. Seasonal analyses showed the expected summer peaks with unexpected off-season outbreaks in 2016, 2017, and 2022–2024. Additionally, EV positivity rates declined significantly during the SARS-CoV-2 pandemic (p = 0.006) followed by a resurgence in 2022–2023. Phylogenic analysis revealed genomic shifts in CVB2, E-5, E-6, E-18, and E-30, whereas CVB5 and E-4 showed no significant variations. CVB2 infections were particularly associated with severe illness in infants, with 10.6% of all sequenced cases requiring ICU admission. These findings demonstrate dynamic changes in EV epidemiology in Israel, including shifts in seasonality, genotype variability, and the impact of SARS-CoV-2 on EV circulation, highlighting the importance of continued molecular surveillance for identifying emerging strains and better understand trends relevant to public health.

Serum Mac-2 binding protein glycosylation isomer (M2BPGi) formation requires sugar chain glycosylation on M2BP. Whether glycated hemoglobin (HbA1c) levels affect M2BPGi in diagnosing hepatitis C virus (HCV)-related hepatic fibrosis remains unclear. We enrolled 2064 patients with available M2BPGi, HbA1c, and valid vibration-controlled transient elastography (VCTE) for fibrosis staging. Associations between M2BPGi and HbA1c levels across fibrosis stages were analyzed using Kruskal–Wallis test and Spearman's correlation. Multivariable linear regression with Akaike information criterion (AIC) identified factors associated with M2BPGi levels. The diagnostic accuracy of M2BPGi in fibrosis staging was evaluated using the areas under receiver operating characteristics (AUROCs) with 95% confidence intervals (CIs). M2BPGi levels showed no correlation with HbA1c categories (< 5.7%, 5.7%–6.4%, 6.5%–6.9%, 7.0%–7.9%, ≥ 8.0%) in fibrosis stages F2 (p = 0.48), F3 (p = 0.63), or F4 (p = 0.22), except in F0–F1 (p < 0.001). Spearman's ρ for F0–F1, F2, F3, and F4 were 0.086 (p = 0.0088), 0.087 (p = 0.057), 0.056 (p = 0.41), and −0.023 (p = 0.62), respectively. Multivariable regression identified fibrosis stage (β for F4, F3, F2 vs. F0–F1 = 4.2724, 1.7623, 0.7223; p < 0.001), age (β = 0.0159; p < 0.001), ALT quotient (β = 0.0094; p < 0.001), steatosis (β = 0.2460; p < 0.001), but not HbA1c, as independent predictors of M2BPGi. The diagnostic accuracy of M2BPGi for fibrosis staging was comparable across HbA1c categories. In conclusion, HbA1c levels do not influence the diagnostic performance of M2BPGi for HCV hepatic fibrosis staging.

HIV controllers (HICs) maintain low-level viremia and normal CD4⁺ T-cell counts without antiviral therapy (ART). This study aimed to characterized immunophenotypic and reservoir dynamics in Chinese HICs, revealing key distinctions from normal progressors (NPs). This prospective single-center study enrolled HICs (ART-free HIV RNA < 2000 copies/mL ≥ 12 months) and age/sex-matched NPs served as controls at PUMCH (2000–2024). Longitudinal immunophenotyping of PBMCs was performed and used PCR to detected total HIV-1 DNA. Among 1792 PLWH, 46 (2.5%) HICs (median age 31 years, 69% male) were enrolled in this study underwent 6 (5–12.5) years follow-up, including 8 elite controllers and 38 viral controllers. Baseline CD4⁺ T-cell counts were 567 (507–757) cells/μL, VL 2.64 (2.35–2.98) log10 copies/mL. Compared with NPs, HICs exhibited higher NK-cell percentage, B-cell percentage, naive CD4⁺ T-cell counts and CD4/CD8 ratio, lower activated CD8⁺ T-cells (CD38⁺orHLA-DR⁺) percentage and total HIV-DNA at baseline (p < 0.05). ART reduced HIV-DNA in NPs but remained higher than HICs (p < 0.05). Five HICs discontinued follow-up, 43.9% (18/41) HICs lost virological control at 6 (5–9) years. HICs experiencing lost virological control exhibited elevated baseline HIV-DNA, activated CD8⁺ T-cells percentage and lower CD4/CD8 ratio. Preceding control loss, declining CD4⁺ T-cell counts (predominantly naive subsets), CD4/CD8 ratios and rising activated CD8⁺ T-cells percentage were observed. Conversely, HICs maintaining virological control retained stable CD38⁺CD8⁺ T-cell percentages, CD4/CD8 ratio and naive CD4⁺ T-cell counts exhibited increased elevated HLA-DR⁺CD8⁺ T-cell percentage. Chinese HICs exhibit different immunophenotypes with reduced HIV-DNA versus NPs. Longitudinal tracking of HIV-1 DNA, activated CD8⁺ T-cell, naïve CD4⁺ T-cell dynamics, and CD4/CD8 stability predict virologic failure in HICs.

The main protease (Mpro) from coronaviruses represents an attractive therapeutic target for antiviral development. The fluorescence resonance energy transfer (FRET) assay is widely used for high-throughput screening (HTS) of Mpro inhibitors, but there has been a significant increase in false positives stemming from flawed assay design in previous studies. Here, we provide an overview of the FRET assay, discuss the key points of this method design, and highlight the corresponding solutions. We hope that this issue should receive increased attention from researchers.