Arbind Kumar

1.0. In the recent decades, the world coped with various waves of severe acute respiratory syndrome (SARS), an emerging infectious viral disease characterized by severe clinical manifestations of t...

Introduction: The aim of this study is to develop a one-step real-time PCR assay for SARS-CoV-2 detection. The study was designed to circumvent the routine RNA isolation step and to optimize a lysis buffer and parameters for direct quantitative PCR. Methodology: A lysis solution was prepared using Tween-20, Triton X-100, EDTA, and Tris buffer (pH 7.4). Various parameters including the use of detergent combinations, V/V ratios and usage of carrier molecules were standardized to achieve the optimal amplification curve and Ct values of SARS-CoV-2 gene for improving the routine diagnostics procedures. Results: Adding carrier molecules [Poly(A), glycogen, and linear polyacrylamide] to the lysis solution significantly improved real-time reverse-transcription PCR (rtRT-PCR) efficacy. Poly(A) was the most effective of all carriers. The diagnostic potential of this Poly(A) solution was demonstrated using 150 patient swabs infected with SARS-CoV-2 and 200 uninfected swab samples, and the sensitivity of the rtRT-PCR diagnostic test was estimated to be 98.6 (95% CI: 96.0, 101.17, p < 0.001) for group 1; Ct ≤ 25 and 87.2 (95% CI: 80.2, 94.0, p < 0.001) for group 2; Ct ≥ 26–30, with excellent accuracy (0.9 < AUC < 1.0), and 100% specificity. Conclusions: Our finding imply that this strategy is feasible, and it may contribute to the development of a rapid, less laborious, and economical rtRT-PCR test for dealing with the SARS-COV-2 disease in the pandemic area.

Aim: To decipher the role of MSMEG_5850 in the physiology of mycobacteria. Methods: MSMEG_5850 was knocked out and RNA sequencing was performed. MSMEG_5850 protein was purified from the Escherichia... Graphical abstract

Background: The global incidence of SARS-CoV-2 infection is now very low. Despite the vaccination drive worldwide, the emergence of new omicron variants and their infection have been reported in a few countries. It is now required to identify potential risks associated with the COVID-19 disease aftermath’s 3rd disease wave. During pandemic stages, the healthcare system bears a significant burden in identifying early risk and providing early management to reduce infection. Methods: In this investigation, an attempt has been made to assess the laboratory risk of SARS-CoV-2 contamination. The samples were collected from the various blocks of the testing centre, including the surface, floor, diagnostic instruments, solutions, and tap water, and then RNA was isolated and in vitro diagnostic RT-qPCR was performed. A total of 316 samples were collected and analysed for the presence of SARS-CoV-2 RNA. Our findings confirmed that only four samples (1.77%) had ORF-1ab and E gene signals, which indicated the presence of SARS-CoV-2 RNA. The CT values for the E gene were 34.52 [IQR: 32.37-36.36] and 35.02 [IQR:34.25-35.76] for the ORF1-ab gene, respectively. Results: All four were taken from the surfaces, indicating that other parts were contamination-free. Very few positive contaminations demonstrated that laboratory sterilising processes are fully functional and effective. Conclusion: This study eventually provided more information about the presence of SARS-CoV-2 RNA traces in the environment aftermath of 3rd disease wave of COVID-19, including diagnostic laboratories.

Introduction: The coronavirus disease 2019 (COVID-19) wave has fluctuated erratically around the globe over the past three years of the pandemic, sometimes declining and at other times surging. The cases of infection in India have remained low, despite the continued surge of Omicron sub-lineages reported in a few countries. In this study, we determined the presence of the circulating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains in the population of Kangra District, Himachal Pradesh, India. Methodology: In vitro diagnostic real-time reverse transcriptase polymerase chain reaction (RT-qPCR) was performed using Tata MD CHECK RT-PCR Omisure kit (Tata Medical and Diagnostics Limited, Maharashtra, India), to detect the presence of Omicron in target samples. A total of 400 samples were analyzed in this study; 200 each for the second and third waves, respectively. The S gene target failure (SG-TF) and S gene mutation amplification (SG-MA) primer-probe sets were used. Results: Our results corroborated that during the third wave, SG-MA amplification was noted, while amplification of SG-TF was not, and vice versa in the case of the second wave, indicating that all the tested patients were infected with the Omicron variant during the third wave, while Omicron was absent during the second wave. Conclusions: This study added more information about the prevalence of Omicron variants during the third wave in the chosen area, and it projected a use of in vitro RT-qPCR method for rapid prospective determination of the prevalence of the variant of concern (VOC) in developing countries with limited sequencing facility.

Background: Highly evolved biocatalysts that can endure harsh environmental conditions during industrial processes are highly desirable. The availability of suitable biocatalysts with high enzyme activity, substrate selectivity, and stability could lower the production costs in the pharmaceutical, chemical, and food industries, resulting in more economical products. Objectives: Naturally evolved enzymes could not be exploited in industrial applications because of their compromised properties. Till date, protein engineering strategies have helped us to improve the desired physical and catalytic properties of enzymes to meet their performance needs in industrial and medical applications. Results: Protein engineering technologies such as directed evolution and rational redesigning are well-suited for improving biocatalytic properties. Each approach has its own set of limitations, and the implementation of techniques is contingent on the availability of prerequisite information about the biocatalyst. Protein structure information is essential for rational design, but no prior structural knowledge is required for directed evolution. Furthermore, semi-rational approaches and enzyme designing are also being used. Considering these facts, this study outlines the various molecular techniques used to improve the physical and catalytic properties of enzymes. It also emphasises the magnitude of strategies used to improve the properties of biocatalysts to meet the needs of industrial processes. Conclusion: Protein engineering frequently employs for improving crucial enzyme characteristics. A semi-rational approach has now emerged as the preferred technology for protein engineering. However, adopting an engineering strategy to achieve the desired characteristic depends on the availability of resources and subject-matter knowledge.

The aim of this study is to develop a one-step real-time PCR assay for SARS-CoV-2 detection. A lysis solution was prepared using Tween-20, Triton X-100, EDTA, and tris buffer (pH 7.4) and various parameters were optimised. Adding carrier molecules [Poly (A), glycogen, and linear polyacrylamide] to the lysis solution significantly improved RT-qPCR efficacy. Poly (A) was the most effective of all carriers. Diagnostic potential of this Poly (A) solution was demonstrated using 150 positives and 200 negative swabs, and the sensitivity of the RT-qPCR diagnostic test was estimated to be 98.6 (95%CI; 96.0, 101.17, p < 0.001) for group 1; Ct ≤ 25 and 87.2 (95%CI; 80.2, 94.0, p < 0.001) for group 2; Ct ≥ 25–30, with excellent accuracy (0.9 < AUC < 1.0), and 100% specificity.

The SARS-CoV-2 outbreak has posed a plethora of problems for the global healthcare system and socioeconomic burden. Despite valiant efforts to contain the COVID-19 outbreak, the situation has deteriorated to the point that there are no viable preventive therapies to treat this disease. The case count has skyrocketed globally due to the newly evolved variants. Despite vaccination drives, the re-occurrence of recent pandemic waves has reinforced the importance of innovation/utilization of immune-booster to achieve appropriate long-term vaccine protection. Plant-derived immuno-adjuvants, which have multifaceted functions, can impede infections by boosting the immune system. Many previous studies have shown that formulation of vaccines using plant-derived adjuvant results in long-lasting immunity may overcome the natural tendency of coronavirus immunity to wane quickly. Plant polysaccharides, glycosides, and glycoprotein extracts have reportedly been utilized as enticing adjuvants in experimental vaccines, such as Advax, Matrix-M, and Mistletoe lectin, which have been shown to be highly immunogenic and safe. When employed in vaccine formulation, Advax and Matrix-M generate long-lasting antibodies, a balanced robust Th1/Th2 cytokine profile, and the stimulation of cytotoxic T cells. Thus, the use of adjuvants derived from plants may increase the effectiveness of vaccines, resulting in the proper immunological response required to combat COVID-19. A few have been widely used in epidemic outbreaks, including SARS and H1N1 influenza, and their use could also improve the efficacy of COVID-19 vaccines. In this review, the immunological adjuvant properties of plant compounds as well as their potential application in anti-COVID-19 therapy are thoroughly discussed.