Assay for Rapid Quantification of Capped and Tailed Intact mRNA

Application technology to fight the COVID-19 pandemic: Lessons learned in Thailand

In recent years, vaccines have played a critical role in protecting against a wide range of diseases, from influenza to polio. As the world continues to grapple with the COVID-19 pandemic, the need for effective vaccines has never been greater. In Pakistan, the situation is no different. According to data from the World Health Organization, Pakistan has seen over 880,000 confirmed cases of COVID-19 and over 18,000 deaths as of January 2021 [1]. With numbers like these, it is clear that the development of vaccines is essential for bringing the pandemic under control and protecting the people of Pakistan.
 One of the primary reasons for the importance of vaccine development in Pakistan is the country's relatively limited resources. While many developed nations have the financial and technological resources to effectively manage the spread of COVID-19 through measures such as widespread testing and contact tracing, Pakistan has fewer resources at its disposal. In such a situation, vaccines provide an important tool for controlling the spread of the virus and protecting the population. Another reason for the need for vaccine development in Pakistan is the country's high population density. With over 220 million people living in a small area, Pakistan has one of the highest population densities in the world [2]. In such a densely populated country, it is especially important to have effective measures in place to prevent the spread of infectious diseases like COVID-19. Vaccines can play a crucial role in this regard by providing immunity to large numbers of people, thereby reducing the overall transmission of the virus.
 There is also the issue of economic impact to consider. The COVID-19 pandemic has had a devastating effect on economies around the world, and Pakistan has been no exception. According to the World Bank, the pandemic is expected to have a significant impact on Pakistan's economy, with GDP growth forecast to fall from 2.4% in 2020 to -0.5% in 2021 [2]. One way to mitigate the economic effects of the pandemic would be to bring it under control as quickly as possible, and vaccines are an essential tool for achieving this.
 In order to address these challenges and improve vaccination rates in Pakistan, it is essential that efforts be made to increase the availability and accessibility of vaccines. This could involve investing in infrastructure and training to ensure that vaccines are properly stored, handled, and administered, as well as implementing outreach programs to educate the public about the importance of vaccination. In addition to these efforts, there is also a need for ongoing vaccine research and development in Pakistan.
 In conclusion, the need for vaccine development in Pakistan is clear. With limited resources, a high population density, and a struggling economy, the country needs effective tools for controlling the spread of COVID-19 and protecting its citizens. Vaccines have the potential to play a crucial role in this regard, and it is important for Pakistan to prioritize their development and distribution. By prioritizing vaccine development and increasing access to vaccines, we can take an important step towards a healthier and more prosperous future for all.

Since the COVID-19 pandemic, it has become very important to teach undergraduate students at all levels, about disease transmission and vaccine biology. Just using lectures and reading assignments on these topics often will not be very effective for the student learning. In resource limited undergraduate teaching environment, it is often not possible to use advanced immunology lab techniques. To combat this problem, a lab-module was incorporated into the undergraduate immunology course. In this module, students will follow the process of vaccine development from the identification of a novel disease through the creation and clinical testing of a vaccine This hands-on module was run over the course of three-week lab period, including important concepts of disease transmission, disease diagnosis, mRNA vaccine development, vaccine efficacy and herd immunity. All the exercises included paper-simulations and simulated samples using sodium hydroxide pellets. Overall concepts of innate and adaptive immunity, and detailed concepts like antigenic epitopes, primary and secondary immune responses, and booster doses are emphasized. Students also learn to design mRNA vaccines using gene sequences of Coronavirus spike protein. They also learn principles of ELISA and phases of clinical trial of a novel vaccine. A survey was conducted at the end of the module to evaluate the student learning and student responses to this module. 90% of the students in class were very pleased with this module and stated that this module helped them consolidate their understanding of vaccines. This module can also be modified to be used in a molecular biology or upper-level microbiology courses, with minimum resources. None

The COVID-19 pandemic is frequently compared to historical influenza outbreaks, yet this analogy often obscures fundamental biological and epidemiological divergences. As the global health community confronts new threats like H5N1 avian influenza, a rigorous retrospective analysis is essential to distinguish the unique trajectory of SARS-CoV-2 from the influenza pandemics of 1957 (H2N2), 1968 (H3N2), and 2009 (H1N1). This narrative review, adhering to best-practice guidance, synthesises literature published up to mid-2025. We performed targeted searches of PubMed, EMBASE, and WHO/CDC archives to compare SARS-CoV-2 critically and Influenza A across virological, clinical, and sociopolitical dimensions. Our analysis reveals that while both pathogens share respiratory transmission, they differ starkly in evolutionary mechanisms and systemic impact. Virologically, SARS-CoV-2 utilises convergent evolution and broad ACE2 tropism to cause multi-organ pathology and long-term sequelae (Long COVID), distinguishing it from the primarily respiratory effects and reassortment-driven shifts of influenza. Epidemiologically, SARS-CoV-2 exhibits a significantly higher basic reproduction number (R 0 ), rising from ~ 2.5 to > 9.5 in Omicron variants, and substantial asymptomatic transmission, necessitating non-pharmaceutical interventions of unprecedented scale compared to the modest measures of past flu pandemics. The public health response also highlighted a divergence in medical innovation: the development of mRNA vaccines within 11 months represented a “quantum leap” in vaccinology compared to the slower, egg-based timelines of 2009. In conclusion, COVID-19 represents a distinct biological and societal entity that defies the historical “flu template.” The transition to endemicity, marked by year-round waves of immune-evasive variants like JN.1, and the current emergence of mammalian H5N1 underscore the need for pathogen-agnostic preparedness. Future global health security depends on leveraging the genomic and technological assets developed during COVID-19 to counter threats that transcend historical expectations.

This paper delves into the application and prospects of bio-antibiotics and mRNA vaccines in preventing and treating bacterial infections and infectious diseases. Against the backdrop of the increasingly serious antibiotic resistance issue, the effectiveness of traditional antibiotic treatments is challenged, while bio-antibiotics and mRNA vaccines, as novel treatment methods, are emerging. Bio-antibiotics, derived from organisms or bioengineered, possess advantages such as antibacterial activity, biocompatibility, low toxicity, and reduced resistance risk, showing broad prospects in treating multi-drug-resistant bacteria infections, special types of infections, and preventive medication. mRNA vaccines, by introducing mRNA encoding pathogen antigens to stimulate the human immune response, are characterized by rapid development, high efficiency, and safety, playing a crucial role in infectious disease prevention and control, especially during the COVID-19 pandemic when the successful development and widespread application of mRNA vaccines significantly reduced the infection rate and mortality. The article also analyzes successful cases of bio-antibiotics and mRNA vaccines in practical applications and the challenges they face, such as extraction and purification techniques, stability, delivery systems, long-term safety, and regulatory policies, and proposes future research directions and prospects. In conclusion, bio-antibiotics and mRNA vaccines, as novel treatment, have great application potential and will bring new hope and opportunities to the global public health field.

Staffing shortages have been widely reported in US nursing homes during the COVID-19 pandemic, but traditional quantitative research analyses have found mixed evidence of staffing shortfalls. To examine whether nursing home administrator perspectives can provide context for conflicting aggregate staffing reports in US nursing homes during the COVID-19 pandemic. In a qualitative study, convergent mixed-methods analysis integrating qualitative and quantitative data sets was used. Semistructured qualitative interviews were conducted between July 14, 2020, and December 16, 2021. Publicly available national Payroll Based Journal data were retrieved from January 1, 2020, to September 30, 2022, on 40 US nursing homes in 8 health care markets that varied by region and nursing home use patterns. Staffing and resident measures were derived from Payroll Based Journal data and compared with national trends for 15 436 US nursing homes. Nursing home administrators were recruited for interviews. Of the 40 administrators who consented to participate, 4 were lost to follow-up. Four repeated, semistructured qualitative interviews with participants were conducted. Interview questions focused on the changes noted during the COVID-19 pandemic in nursing homes. Thematic description of nursing home administrator compensatory strategies to provide context for quantitative analyses on nursing home staffing levels during the COVID-19 pandemic. A total of 156 interviews were completed with 40 nursing home administrators. Administrators reported experiencing staff shortages during the COVID-19 pandemic and using compensatory strategies, such as overtime, cross-training, staff-to-resident ratio adjustments, use of agency staff, and curtailing admissions, to maintain operations and comply with minimum staffing regulations. Payroll Based Journal data measures graphed from January 1, 2020, to September 30, 2022, supported administrator reports showing that study facilities had reductions in staff hours, increased use of agency staff, and decreased resident census. Findings were similar to national trends. In this qualitative, convergent mixed-methods study, nursing home administrators reported the major staffing strain they experienced at their facilities and the strategies they used to offset staffing shortages. Their experiences provide context to quantitative analyses on aggregate nursing home census data. The short-term compensatory measures administrators used to comply with regulations and maintain operations may be detrimental to the long-term stability of this workforce.

With the rapid success in the development of mRNA vaccines against COVID-19 and with a number of mRNA-based drugs ahead in the pipelines, mRNA has catapulted to the forefront of drug research, demonstrating its substantial effectiveness against a broad range of diseases. As the recent global pandemic gradually fades, we cannot stop thinking about what the world has gained: the realization and validation of the power of mRNA in modern medicine. A significant amount of research has now been concentrated on developing mRNA drugs and vaccine platforms against infectious and immune diseases, cancer, and other debilitating diseases and has demonstrated encouraging results. Here, based on the CAS Content Collection, we provide a landscape view of the current state, outline trends in the research and development of mRNA therapeutics and vaccines, and highlight some notable patents focusing on mRNA therapeutics, vaccines, and delivery systems. Analysis of diseases disclosed in patents also reveals highly investigated diseases for treatments with these medicines. Finally, we provide information about mRNA therapeutics and vaccines in clinical trials. We hope this Review will be useful for understanding the current knowledge in the field of mRNA medicines and will assist in efforts to solve its remaining challenges and revolutionize the treatment of human diseases.

The rapid evolution of pathogen threats and pandemic risks has intensified the demand for adaptable vaccine platforms with rapid design-to-clinical timelines. The success of BNT162b2 and mRNA-1273 in controlling COVID-19 has led to unprecedented changes in vaccine development. Compared with traditional platforms, mRNA vaccines offer distinct advantages: avoiding genomic insertion risks of DNA vaccines; facilitating rapid, scalable production via in vitro transcription; and enabling new modular antigen design against evolving pathogens. However, the development of mRNA vaccines for infectious diseases still faces challenges, including structural stability, translation efficiency, and targeted delivery. This review presents cutting-edge strategies for designing antigens, optimizing mRNA structural elements, and optimizing delivery systems to target infectious pathogens. This study provides a detailed explanation of antigen optimization strategies for different types of mRNA vaccines and explores the potential applications of broad-spectrum and combination mRNA vaccines in human and animal pathogens. Finally, we address the current challenges and future prospects of mRNA vaccine technology, aiming to inform and accelerate the development of next-generation vaccines against emerging pathogens through the integration of these innovations.

The relentless global burden of Tuberculosis (TB), causing over 10.7 million new cases and 1.25 million deaths in 2024, demands urgent innovation [1]. The limited and variable efficacy (0-80%) of the Bacille Calmette-Guérin (BCG) vaccine in adults, a challenge starkly highlighted during the COVID-19 pandemic, underscores the critical need for novel prophylactic and therapeutic strategies [2][3][4][5]. This third volume of our Research Topic, "Community Series in Research Advances of Tuberculosis Vaccine and its Implication on COVID-19," collates pioneering work that bridges immunology, nanotechnology, and clinical research. The eight articles herein not only provide deep insights into TB immunopathogenesis but also present tangible advances in vaccine design, diagnostic precision, and patient stratification, with broader implications for managing intracellular infections.This collection comprises eight articles (4 Research Articles, 2 Reviews, 2 Corrections) that collectively address critical bottlenecks in TB immunology, vaccine development and immunodiagnostics (Table 1). • Induced a strong DTH response in guinea pigs (induration of 18.3 ± 1.8 mm at 10μg/ml) with no cross-reactivity to BCG.• Preclinical assessment confirmed excellent safety and specificity, supporting its potential for clinical trials.Frontiers in Immunology • Identified a 3-gene cuproptosis-related signature (ASPHD2, GK, GCH1) for PTB diagnosis with high accuracy (AUCs: 0.981, 0.928, 0.937).• The signature was linked to a significantly altered immune microenvironment in PTB patients.• GCH1 and GK expression significantly decreased after one month of treatment (p<0.05),suggesting their utility as therapy monitoring biomarkers.Research Article Masoud Mortezazadeh et al.• LTBI prevalence was not significantly higher in cancer patients (27.1%) vs. controls (20.7%, P=0.176).• However, LTBI was a strong predictor of mortality in cancer patients (OR=3.28, P<0.001).• Each 1mm increase in TST induration was associated with a 6% increased risk of death (HR=1.06, P=0.001).5 Review Aishwarya Shaji et al.• Systematically reviewed the TB vaccine pipeline, noting the variable efficacy of BCG (0-80%).• Highlighted promising candidates like M72/AS01E, which showed 54% efficacy against MTB infection in adults.• Emphasized that host genetics (e.g., IFN-γ +874 T/A polymorphism) are a core determinant of vaccine efficacy, advocating for precision vaccinology. • Highlighted that MTB-specific IgM in BALF was the strongest correlate of reduced bacterial burden in primates.• Argued that future vaccines must be designed to elicit a precisely tuned humoral immune response. • Corrected errors in the labeling of figures in the supplementary materials.• Confirmed that the corrections do not affect any experimental data, statistical results, or the main conclusions of the study. • Restored the abstract to the authors' originally submitted version after a production error.• Clarified that the correction is textual only and does not alter any data or scientific conclusions presented in the article.The collective findings presented in this Research Topic create a powerful, synergistic narrative. They move the field forward not in isolated increments, but through a convergent evolution of ideas. However, the review by Huoming Li and Hao Li compellingly argues that this is an incomplete picture. The next frontier may lie in designing vaccines that not only elicit potent T-cell immunity but also guide the development of a protective antibody profile [8][9][10][11][12][13],perhaps using the very targeting technologies showcased in this topic. This integrated approach-orchestrating both arms of the adaptive immune system-is a cornerstone of modern vaccinology, as learned from COVID-19. The collective works in this Research Topic vividly illustrate a field in rapid and sophisticated evolution. From the nano-engineering of targeted immune stimulants and the computational discovery of diagnostic biomarkers, to the refined understanding of immune correlates and the translation of epidemiological data into clinical practice, this volume showcases a multi-faceted attack on TB. The integration of key quantitative findings-from vaccine efficacy rates and diagnostic AUC values to relative risk metricsprovides a solid evidence base for future research. These advances, particularly the lessons in leveraging innate immunity and understanding host-specific responses, resonate deeply with the broader vaccinology landscape, including the fight against COVID-19. As we move forward, the continued synergy between disruptive technology, basic immunology, and precision medicine will be paramount in achieving the ultimate goal: ending the TB epidemic.

The development of severe multidrug-resistant bacterial infections has recently intensified because of the COVID-19 pandemic. According to the guidelines issued by the World Health Organization (WHO), routine antibiotic administration is not recommended for patients with supposed or confirmed mild SARS-CoV-2 infection or pneumonia, unless bacterial infection is clinically suspected. However, recent studies have pointed out that the proportion of non-essential antibiotic use in patients infected with SARS-CoV-2 remains high. Therefore, the silent pandemic of antibiotic resistance remains a pressing issue regardless of the present threats presented by the COVID-19 pandemic. To prevent or delay entry into the postulated post-antibiotic era, the long-term advocacy for the rational use of antibiotics, the optimization of infection control procedures, and the development of new antibacterial agents and vaccines should be underscored as vital practices of the antibacterial toolbox. Recently, the development of vaccines and monoclonal antibodies has gradually received attention following the advancement of biotechnology as well as enhanced drug discovery and development in cancer research. Although decent progress has been made in laboratory-based research and promising results have been obtained following clinical trials of some of these products, challenges still exist in their widespread clinical applications. This article describes the current advantages of antibacterial monoclonal antibodies, the development of associated clinical trials, and some perceived future perspectives and challenges. Further, we anticipate the development of more therapeutic agents to combat drug-resistant bacterial infections as well as to increase the resilience of current or novel agents/strategies.

Bluetooth Low Energy (BLE) based Wireless Indoor Localization System (WILS) with high localization accuracy and high localization precision is a key requirement in enabling the Internet of Things (IoT) in today’s applications. In this paper, we investigated the effect of BLE signal variations on indoor localization caused by the change in BLE transmission power levels. This issue is not often discussed as most of the works on localization algorithms use the highest power levels but has important practical implications for energy efficiency, e.g., if a designer would like to trade-off localization performance and node lifetime. To analyze the impact, we used the established trilateration based localization model with two methods i.e., Centroid Approximation (CA) and Minimum Mean Square Error (MMSE). We observed that trilateration based localization with MMSE method outperforms the CA method. We further investigated the use of two filters i.e., Low Pass Filter (LPF) and Kalman Filter (KF) and evaluated their effects in terms of mitigating the random variations from BLE signal. In comparison to non-filter based approach, we observed a great improvement in localization accuracy and localization precision with a filter-based approach. Furthermore, in comparison to LPF based trilateration localization with CA, the performance of a KF based trilateration localization with MMSE is far better. An average of 1 m improvement in localization accuracy and approximately 50% improvement in localization precision is observed by using KF in trilateration based localization model with the MMSE method. In conclusion, with KF in trilateration based localization model with MMSE method effectively eliminates random variations in BLE RSS with multiple transmission power levels and thus results in a BLE based WILS with high accuracy and high precision.

Nucleic acid vaccines have attracted enormous attention for resolving the limitations of conventional vaccines using live attenuated viruses. Because nucleic acid vaccines can be produced rapidly in response to the emergence of new virus strains, they are more appropriate for the control of urgent epidemic and pandemic issues. In particular, messenger RNA (mRNA) vaccines have evolved as a new type of nucleic acid vaccines in accordance with their superior protein expression and a lack of mutagenesis as compared with DNA vaccines. Using mRNA vaccines, large amounts of target proteins can be expressed in immune cells for efficient immunization. For instance, antigen-specific vaccination is a feasible option involving the expression of specific antigens in antigen-presenting cells. Immunological reactions are modulated by expressing several proteins associated with stimulation or maturation of immune cells. In addition, mRNA vaccines can stimulate innate immunity through specific recognition by pattern recognition receptors. On the basis of these remarkable properties, mRNA vaccines have been used for prophylactic and therapeutic applications. This review highlights the role of mRNA vaccines as prophylactic vaccines for prevention of future infections and as therapeutic vaccines for cancer immunotherapy. In addition to the conventional type of mRNA vaccines, RNA replicons (self-amplifying mRNA vaccines) will be described.

mRNA vaccines for infectious diseases prevent diseases by stimulating the body to produce specific immune responses through mRNA molecules encoding pathogenic proteins. Compared to traditional vaccines (e.g., inactivated, live attenuated, subunit, recombinant protein and viral vectors), mRNA vaccines offer several advantages including high safety, potent efficacy, scalable large-scale production, and cost-effectiveness. mRNA vaccines have demonstrated significant potential in combating infectious diseases since their inception. In particular, during the 2019 Coronavirus Disease (COVID-19) pandemic, the mRNA vaccines delivered with lipid nanoparticles (LNPs) have been developed by BioNTech and Moderna, their exceptional protective efficacy and extensive clinical application further proved the rapid responsiveness of mRNA vaccines in addressing emerging infectious diseases. This success has brought mRNA vaccines back into the spotlight of the scientific community. This article reviews the molecular biological basis, delivery systems, and immune mechanisms of mRNA vaccines, as well as the progress of research and clinical trials related to mRNA vaccines targeting the COVID-19 virus, influenza virus, rabies virus, Zika virus, human immunodeficiency virus, and Mycobacterium tuberculosis (M.tb), while also discussing the current challenges faced in the application of mRNA vaccines. These discussions provide a theoretical foundation and practical guidance for the future development of mRNA vaccines targeting bacterial infectious diseases such as tuberculosis (TB).Supplementary InformationThe online version contains supplementary material available at 10.1186/s40001-025-03060-x.

A highly sensitive nanopore platform for measuring RNase A activity

World AIDS Day 2022: A bittersweet commemoration of the global HIV/AIDS response.