The enduring SARS-CoV-2 pandemic necessitates robust tools for severity assessment. This study, conducted at Islamabad Diagnostic Center across Pakistan from January 2021 to August 2022, aimed to investigate hematological abnormalities among suspected SARS-CoV-2 subjects. Initial enrollment included 130,347 cases, with 53,078 confirmed positive and 77,269 negative. An additional 11,786 samples expanded the dataset to 142,133. The Omicron and Centaurus variants, in confirmed positive patients, exhibited a slightly higher frequency of hematological abnormalities (30.42%) than negative participants (27.01%). Notably, lymphocyte count reduction (40.95%) suggested its potential as an alternative diagnostic parameter for clinical COVID-19. Decreased levels of NA (37.99%), HGB (26.17%), MCV (20.60%), PLT (6.15%), and ALB (2.28%) were observed. Abnormally elevated NEU, CR, MONO, RBCs, WBC, and EOS levels affected 26.00%, 24.28%, 30.79%, 22.02%, 6.28%, and 5.53% of subjects, respectively. Comparatively, positive patients exhibited higher abnormal blood parameters—LYMP count (57.40%), NEU count (46.08%), EOS count (62.48%), MONO count (31.61%), RBC count (30.32%), ALC count (43.60%), CR count (30.91%), NA count (40.53%), CRP count (68.46%), and DD (63.08%) than negative counterparts. The study underscores lymphocytopenia’s potential as a cost-effective, early diagnostic biomarker for clinical COVID-19, preceding real-time PCR diagnosis. This supports its consideration in resource-limited settings for strategic screening and policy-making in the ongoing SARS-CoV-2 battle.

Background: Hepatitis B virus (HBV) establishes persistent infection through the formation of a covalently closed circular DNA (cccDNA) minichromosome, which enables immune evasion and sustained viral replication. While SIRT1, IFI16, and STING are known to modulate antiviral pathways, their cooperative role in regulating HBV transcription and host immunity remains poorly defined. Methods: We investigated the functional interplay between SIRT1, IFI16, and STING using confocal microscopy, chromatin immunoprecipitation (ChIP), gene silencing, and quantitative PCR in HBV-replicating cell models. Interactions with HBV cccDNA and effects on viral transcription and interferon-stimulated gene (ISG) expression were analyzed. Northern blotting and RT-qPCR were used to evaluate HBV RNA output, including total HBV RNA and 3.5 kb RNA, in order to provide both transcript-pattern and quantitative assessment of transcriptional changes. Results: SIRT1 physically interacted with IFI16, with increased colocalization during HBV replication. Both proteins were recruited to HBV cccDNA, promoting transcriptionally active chromatin. Silencing SIRT1 or IFI16 disrupted cccDNA binding and reduced viral RNA levels. Although STING was not cccDNA-associated, its antiviral activity via ISG induction was dependent on IFI16 knockdown produced the strongest reduction in both total HBV RNA and 3.5 kb RNA, supporting a cooperative role for these factors in sustaining HBV transcriptional output. Although STING was not cccDNA-associated, its antiviral activity via ISG induction was dependent on IFI16. Triple knockdown of SIRT1, IFI16, and STING showed the strongest suppression of ISG expression. Conclusion: SIRT1 and IFI16 enhance HBV gene expression through epigenetic regulation of cccDNA, while also supporting STING-mediated antiviral responses. These findings reveal a dual regulatory axis governing HBV persistence and identify potential targets for antiviral therapy.