A new generation of molecular tests has revolutionized laboratory medicine through allowing for quick, sensitive, and precise identification of the pathogen; genetic mutation or alteration; and disease-related bio-markers. Of all the molecular techniques that exist today, PCR is still the most versatile, and clinically applicable technique available. This paper reviews the development of PCR technology from traditional amplification to advanced platforms including qPCR, dPCR, RT-PCR, and multiplex PCR. Each of the advanced platforms reviewed will be examined for their contributions to both the accurate detection, and quantification of nucleic acids along with the ability to perform high throughput analyses. Additionally, this paper also discusses how PCR technology can be integrated with additional technologies such as NGS, CRISPR-based diagnostic systems, microarrays, and epigenetic profiling to provide a larger view of disease states than was previously possible. Specifically, the focus of this paper will include many clinically relevant biomarkers and molecular targets used in cancer research including EGFR, KRAS, BRAF, BCR-ABL1, circulating tumor DNA (ctDNA), and methylated SEPT9. These biomarkers and molecular targets have greatly enhanced early diagnosis, prognostication, treatment monitoring and precision medicine in cancer research and other areas. The use of PCR-based technologies in diagnosing infectious diseases, tracking AMR, detecting heritable disorders, studying microbial communities, and MRD monitoring were also discussed. Further discussion included current trends in integrating multiple omics data types into a single platform using AI assisted data analysis tools. Liquid biopsy technologies and point-of-care molecular test platforms were also discussed. Although there are continuing challenges related to costs associated with developing these technologies, establishing standardized protocols for each technology type developed, having skilled technicians develop and validate these technologies, and managing large amounts of data generated when using these technologies; continued advances are providing greater access to these technologies to clinicians and ultimately patients.
M. Saleem, Taimoor Riaz, Fatima Saleem et al.· Clinica chimica acta; intern...· 0 citations
Drought stress restricts growth and productivity in grain legumes, yet the integrative mechanisms underlying silicon-mediated drought tolerance in Vigna mungo (mash bean) remain poorly understood. The present study aimed to investigate the role of silicon in mitigating drought-induced damage in two mash bean cultivars. To evaluate this effect, plants were subjected to different drought regimes with and without silicon. Data on growth traits, biomass accumulation, reproductive attributes, photosynthetic gas exchange parameters, water relations, antioxidant enzyme activities, phenolic compounds, nutrient dynamics, and expression of drought-responsive genes were collected and statistically analyzed. Multivariate analyses, including PCA and structural equation modeling (SEM), were used to elucidate relationships among measured variables. Drought significantly reduced plant growth, biomass production, and reproductive traits, accompanied by marked declines in net assimilation rate (up to 35-40%), stomatal conductance, and transpiration under 25% field capacity (FC). In contrast, drought increased WUE (49.7-56.2 µmol CO₂ mmol⁻1 H₂O), antioxidant enzyme activities, and phenolic accumulation. Silicon supplementation substantially improved plant growth, restored photosynthetic performance, and enhanced antioxidant defense under drought conditions. Si-treated plants exhibited increased activities of SOD, CAT, and APX, higher phenolic content, improved water status, and upregulated expression of DREB2A, PIP2-1, and TIP4-1 compared with non-supplemented plants. Multivariate analyses clearly separated drought and Si treatments, while SEM revealed strong relationships linking gene expression, biochemical defense systems, physiological processes, and plant growth responses. In conclusion, silicon supplementation significantly enhances drought tolerance in mash bean by improving photosynthetic efficiency, antioxidant capacity, and molecular regulation of water transport.
Z. Ali, Syed Riaz Ahmed, I. Ijaz et al.· Scientific Reports· 2 citations
This review provides a comprehensive synthesis of a recent advances in CRISPR–Cas technologies and their strategic applications in crop genetics and hybrid breeding, and showcases how these technologies accelerate hybrid breeding by engineering male sterility systems, fixing heterosis, and generating high-throughput mutant libraries for trait discovery.
Syed Riaz Ahmed, Jahangir Khan, I. Ijaz et al.· Frontiers in Plant Science· 0 citations