Currently, there are no specific therapeutics for flavivirus infections, including dengue virus (DENV) and Zika virus (ZIKV). In this study, we evaluated extracts from the plants Hedyotis diffusa (HD) and Artemisia capillaris (AC) to determine the antiviral activity against DENV, ZIKV, and Japanese encephalitis virus (JEV). HD and AC demonstrated inhibitory activity against JEV, ZIKV, and DENV replication and reduced viral RNA levels in a dose–responsive manner, with non-cytotoxic concentration ranging from 0.1 to 10 mg/mL. HD and AC had low cytotoxicity to Vero cells, with CC₅₀ values of 33.7 ± 1.6 and 30.3 ± 1.7 mg/mL (mean ± SD), respectively. The anti-flavivirus activity of HD and AC was also consistent in human cell lines, including human glioblastoma (T98G), human chronic myeloid leukemia (K562), and human embryonic kidney (HEK-293T) cells. Viral-infected, HD-treated cells demonstrated downregulation of cytokines including CCR1, CCL26, CCL15, CCL5, IL21, and IL17C. In contrast, CCR1, CCL26, and AIMP1 were elevated following AC treatment in viral-infected cells. Overall, HD and AC plant extracts demonstrated flavivirus replication inhibitory activity, and together with immunoregulatory cytokine signatures, these results suggest that HD and AC possess bioactive compounds that may further be refined as promising candidates for clinical applications.

The RT-qPCR method remains the gold standard and first-line diagnostic method for the detection of SARS-CoV-2 and flaviviruses, especially in the early stage of viral infection. Rapid and accurate viral detection is a starting point in the containment of the COVID-19 pandemic and flavivirus outbreaks. However, the shortage of diagnostic reagents and supplies, especially in resource-limited countries that experience co-circulation of SARS-CoV-2 and flaviviruses, are limitations that may result in lesser availability of RT-qPCR-based diagnostic tests. In this study, the utility of RNA-free extraction methods was assessed for the direct detection of SARS-CoV-2 and DENV-2 in heat-inactivated or chemical-inactivated samples. The findings demonstrate that direct real-time RT-qPCR is a feasible option in comparison to conventional real-time RT-qPCR based on viral genome extraction-based methods. The utility of heat-inactivation and direct real-time RT-qPCR for SARS-CoV-2, DENV-2 viral RNA detection was demonstrated by using clinical samples of SARS-CoV-2 and DENV-2 and spiked cell culture samples of SARS-CoV-2 and DENV-2. This study provides a simple alternative workflow for flavivirus and SARS-CoV-2 detection that includes heat inactivation and viral RNA extraction-free protocols, with aims to reduce the risk of exposure during processing of SARS-CoV-2 biological specimens and to overcome the supply-chain bottleneck, particularly in resource limited settings with flavivirus co-circulation.

Tuberculosis (TB) and non-tuberculous mycobacterial (NTM) detections pose overlapping diagnostic and therapeutic challenges, particularly in high-burden re

Non-tuberculous mycobacteria (NTM) pose diagnostic and therapeutic challenges in tuberculosis (TB)-endemic regions like Jiangxi Province, China, due to cli

Accurate diagnosis of paucibacillary and extrapulmonary tuberculosis remains a major global challenge, as conventional pathogen-focused methods often lack sensitivity. This study aimed to evaluate whether combining host-derived immune biomarkers could improve diagnostic accuracy. In a retrospective diagnostic accuracy study of 605 participants, nucleic acid amplification tests served as the reference standard. We assessed the performance of adenosine deaminase, interferon-gamma, and interleukin-6 individually and in combination using receiver operating characteristic analysis. Serial (rule-in) and parallel (rule-out) testing algorithms were compared to define strategies for confirmation or exclusion of disease. Here we show that while interferon-gamma was the strongest single discriminator, combinations of biomarkers significantly outperform any single marker. The dual-panel of interferon-gamma and interleukin-6 achieves the highest overall accuracy. Serial testing of adenosine deaminase and interferon-gamma maximizes specificity to 97.4% (95% CI: 95.6-98.7%), providing a reliable rule-in tool. Conversely, parallel testing of all three biomarkers achieves 94.9% (95% CI: 95.6-98.7%)sensitivity, enabling an effective rule-out strategy. The integration of complementary host biomarkers overcomes the inherent limitations of single analyte tests. By enabling adaptable, context-specific algorithms tailored for either high-specificity confirmation or high-sensitivity screening, this approach provides a scalable and cost-effective diagnostic approach, tailored for resource-constrained settings.

The long-term success of antiretroviral therapy (ART) is increasingly jeopardized by the emergence of HIV-1 drug resistance (HIVDR). Characterizing the res