Identification, prioritization, and evaluation of RlpA protein as a target against multidrug-resistant Pseudomonas aeruginosa.

According to the World Health Organization, infectious diseases, particularly those caused by multidrug-resistant bacteria (MDR), are projected to claim the lives of 15 million people by 2050. Septicemia carries a higher morbidity and mortality rate than infections caused by susceptible Pseudomonas aeruginosa, and MDR-mediated ocular infections can lead to impaired vision and blindness. To identify and develop a potential drug against MDR P. aeruginosa, we employed in silico reverse genetics-based target mining, drug prioritization, and evaluation. Rare Lipoprotein A (RlpA) was selected as the target protein, and its crystal structure was geometrically optimized. Molecular docking and virtual screening analyses revealed that RlpA exhibits strong binding affinity with 11 compounds. Among these, 3-chlorophthalic acid was evaluated, and subsequent in vitro assays demonstrated significant anti-Pseudomonas activity with negligible cytotoxicity. The compound was further evaluated against both drug-susceptible and MDR P. aeruginosa strains in vitro, with cytotoxicity assessed using an MTT assay. The study demonstrated that 3-chlorophthalic acid exhibits potent anti-Pseudomonas activity with minimal toxicity to host cells. Consequently, this compound emerges as a promising candidate against MDR P. aeruginosa, warranting further investigation.

Unveiling the ocular battlefield: Insights into Pseudomonas aeruginosa virulence factors and their implications for multidrug resistance.

The research investigates the virulence factors of Pseudomonas aeruginosa (P. aeruginosa), a pathogen known for its ability to cause human infections by releasing various exoenzymes and virulence factors. Particularly relevant in ocular infections, where tissue degeneration can occur, even after bacterial growth has ceased due to the potential role of secreted proteins/enzymes. Clinical isolates of P. aeruginosa, both ocular (146) and non-ocular (54), were examined to determine the frequency and mechanism of virulence factors. Phenotypic characterization revealed the production of alginate, biofilm, phospholipase C, and alkaline protease, while genotypic testing using internal uniplex PCR identified the presence of Exo U, S, T, Y, and LasB genes. Results showed a significant prevalence of Exo U and Y genes in ocular isolates, a finding unique to Indian studies. Additionally, the study noted that ocular isolates often contained all four secretomes, suggesting a potential link between these factors and ocular infections. These findings contribute to understanding the pathogenesis of P. aeruginosa infections, particularly in ocular contexts, and highlights the importance of comprehensive virulence factor analysis in clinical settings.

Exploring recombinant secretory proteins from Mycobacterium tuberculosis to develop a serological platform for tuberculosis diagnosis.

The lack of a sensitive diagnostic tool for tuberculosis (TB) is the main reason for increasing cause of death in many developing countries. The routine diagnostic tests are either time-consuming or equivocal in terms of results. Hence, there is a need for quicker and accurate diagnostic tests. Certain studies have documented the usage of proteins secreted by Mycobacterium tuberculosis (MTB) in developing a sensitive tool for diagnosing TB. The study aimed to employ PPE41, MPT53, LPQH, CFP10, ESAT6 and TB18.5 proteins and analyze their usage as early diagnostic markers. The proteins were cloned, expressed, purified and applied in ELISA platforms in separate as well as combined systems to assess their early diagnostic features. The results of our study revealed that a cocktail of all six antigen combinations was identified in the maximum number of TB cases. Thus, proteins such as PPE41, MPT53, LPQH, CFP10, ESAT6, and TB18.5 incorporated detection tools could be optimized for an improvised early detection of MTB infections. Moreover, the results suggested that 95.7 % of the MTB-positive serum samples reacted with all the selected antigens of Mycobacterium tuberculosis, while the control serum samples did not react with those antigens. The hexavalent antigen system yielded a novel ELISA platform for better diagnosing MTB infections. Our study yielded a novel technology to diagnose TB, which warrants testing in clinical settings.

A comparative study on the efficiency of commercial reverse transcriptase-Polymerase chain reaction kits for the detection of severe acute respiratory syndrome coronavirus 2 infections.

Background: Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) kits have been reliably employed for the diagnosis of coronavirus disease 2019 (COVID-19) by the detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA since the beginning of the disease outbreak. In consideration of reliable diagnosis, apart from RT-PCR, the isothermal nucleic acid amplification-based point-of-care automated kits have also been tagged as a simpler and rapid alternative to the conventional techniques. Currently, the availability of a better diagnostic method for COVID-19 when compared to RT-PCR is nil. The most important step in the detection of SARS-CoV-2 in a RT-PCR diagnostic laboratory is to identify and employ RT-PCR kits with higher sensitivity as well as specificity. Objectives: This study aimed to study commercially available RT-PCR kits for the detection of SARS-CoV-2 infections. Methods: The performance of seven different RT-PCR kits from different manufacturers used for diagnosis of COVID-19 in Govt Theni Medical College and Hospital, Theni, Tamil Nadu were analysed. Nasopharyngeal and oropharyngeal swabs were collected from patients and subjected to RT-PCR using these kits. Results and Conclusion: The sensitivities and batch effects of the assessed kits were slightly different for different targets, for SARS-CoV-2 detection in nasopharyngeal swab specimens. Examination of COVID-19 kits should be done using currently employed kits in routine diagnosis for better efficiency.