Tackling hepatitis B Virus with CRISPR/Cas9: advances, challenges, and delivery strategies.

Hepatitis B virus (HBV) infection remains a significant global health challenge, with chronic HBV leading to severe liver diseases, including cirrhosis and hepatocellular carcinoma. Current treatments often fail to eradicate the virus, highlighting the need for innovative therapeutic strategies. The CRISPR/Cas9 system has emerged as a dynamic tool for precise genome editing and presents a promising approach to targeting and eliminating HBV infection. This review provides a comprehensive overview of the advances, challenges, and delivery strategies associated with CRISPR/Cas9-based therapies for HBV. We begin by elucidating the mechanism of the CRISPR/Cas9 system and then explore HBV pathogenesis, focusing on the role of covalently closed circular DNA (cccDNA) and integrated HBV DNA in maintaining chronic infection. CRISPR/Cas9 can disrupt these key viral reservoirs, which are critical for persistent HBV replication and associated liver damage. The application of CRISPR/Cas9 in HBV treatment faces significant challenges, such as off-target effects, delivery efficiency, and immune responses. These challenges are addressed by examining current approaches to enhance the specificity, safety, and efficacy of CRISPR/Cas9. A future perspective on the development and clinical translation of CRISPR/Cas9 therapies for HBV is provided, emphasizing the requirement for further research to improve delivery methods and ensure durable safety and effectiveness. This review underscores the transformative potential of CRISPR/Cas9 in combating HBV and sets the stage for future breakthroughs in the field.

A study of analysis on prevalence, serological marker and prognosis of tuberculosis in tertiary care hospital.

BACKGROUND: Tuberculosis is an infectious disease responsible for a significant cause of ill health. According to the WHO global tuberculosis report 2021. 9.9 million cases fell sick with TB in 2020. Significantly, the prevalence of tuberculosis in India is 25%. OBJECTIVE: To analyze the prevalence of tuberculosis in the suburban areas of the metropolitan city in South India. To analyze the serological marker and prognosis of tuberculosis among males and females. To determine the importance of molecular testing - PCR confirmation on TB after AFB smear. METHODS: A retrospective study to analyze 462 patients enrolled by the respiratory medicine department on suspecting pulmonary- 356 (M-264 & F-92) and extra-pulmonary-106 (M-73&F-33) patients and diagnosed Zhiel-Neelsen staining, Mantoux test, Chip-based RT-PCR test, Erythrocyte sedimentation rate, and analyzed serological test such as C-Reactive Protein, Chemiluminescence immune assay. RESULTS: 23 patients were positive in Ziehl-Neelsen staining, 65 were positive in molecular True-Nat PCR test, Mantoux skin test induration in 10 patients, 98 TB Positive patients examined in the serological analysis, 1 & 3 patients reacted in HIV/HBsAg, and HBsAg test respectively, by chemiluminescence immunoassay, 8 PTB and 4 EPTB and 47 non-TB patients were positive in C-reactive protein, 46 TB and 94 non-TB patients detected abnormal values out of these 160 patients in ESR test. CONCLUSION: The Prevalence of tuberculosis is significantly rising, especially in the middle-aged population. The rapid molecular diagnostics to detect TB are highly sensitive and specific. Serological markers are essential for the analysis of disease prognosis and need to focus on the guidance of DOTS and RNTCP to End TB.

Correlation of phenotypic and genotypic virulence markers, antimicrobial susceptibility pattern, and outcome of Pseudomonas aeruginosa sepsis infection.

INTRODUCTION: Pseudomonas aeruginosa (PA) possesses several virulence genes that enable them to evade the immune system and to cause injury in the host tissue. However, the number of studies that characterized the virulence genes profile in PA sepsis is limited. AIM: The main objective of this study was to identify and characterize virulence genes in PA causing sepsis, as well as investigate the relationship between virulence genes, antimicrobial susceptibility patterns, and infection outcomes. METHODOLOGY: A prospective study, conducted between October 2020-October 2021, isolates were recovered from blood samples and identified using standard microbiological procedures. Phenotypic techniques were used to screen for capsule, siderophore production, biofilm formation, serum resistance, hemolysin production, and protease. Molecular techniques were performed to screen for alginate D (alg D), exoenzyme S (Exo S), exotoxin A (tox A), phospholipase H (plc H), phospholipase N (plc N), and elastase B (las B). Kirby-Bauer disc diffusion method was used to determine the antimicrobial susceptibility pattern of isolates, which was then interpreted according to the CLSI 2021 guidelines. RESULTS: Out of the n = 215 Gram-negative bacteria recovered from sepsis patients during our study, n = 20 were Pseudomonas aeruginosa. PA isolates were susceptible to all antibiotics tested except for 3 of the isolates that were resistant to gentamycin, 2 to imipenem, and 1 to ceftazidime, cefepime, meropenem, tobramycin, and amikacin. The most prevalent virulence genes present were capsule (100%), siderophore production (100%), alg D (100%), Las B (100%), and Tox A (100%). CONCLUSION: Our study found that PA causing sepsis harbours a high level of virulence genes. However, the high presence of virulence factors was not statistically associated with antimicrobial susceptibility, as most isolates in our study were susceptible to the antibiotics tested.