Brahma, U. | bioRxiv Science

bioRxiv2025

Proteomic Insights into Strong and Weak Biofilm Formation in Acinetobacter baumannii for Potential Therapeutic Targets

Acinetobacter baumannii is notable for its biofilm-forming abilities, which aid in its tolerance to antibiotics, adding to antimicrobial resistance. The clinical isolates present varied biofilm-forming capacity; hence, understanding the molecular determinants that result in strong biofilm development is crucial for drug target identification. This study is the first of its kind to compare proteome profiling of strong and weak biofilm-forming A. baumannii clinical isolates. Comparative proteomic profiling revealed 42 differentially regulated proteins. It was observed that in strong biofilm forming isolate NlpA, uL16, DNA gyrase B, acetyl-CoA carboxylase, and purl etc. were upregulated highlighting a dynamic reprogramming of cellular functions that promotes biofilm formation, stress adaptation, and immune evasion. In contrast, EF-Tu, ribosome hibernation factors, and T6SS components were downregulated, suggesting a lack of biosynthesis and stress adaptability. These findings suggest a metabolic downshift and a possible energy conservation mechanism under conditions less favorable for strong biofilm development. Additionally, several uncharacterized proteins were identified, highlighting potential novel factors in biofilm regulation and virulence that warrant further investigation. The proteomics data correlated with qPCR findings, providing support for the unknown regulators of biofilm formation that were identified in this study. Key proteins such as nlpA, 6,7-dimethyl-8-ribityllumazine synthase and DNA gyrase B emerged as potential therapeutic targets. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/680171v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@1c9b95aorg.highwire.dtl.DTLVardef@a7e781org.highwire.dtl.DTLVardef@14fe64aorg.highwire.dtl.DTLVardef@9804f2_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOGraphical AbstractC_FLOATNO C_FIG

microbiology↗

bioRxiv2025

A novel approach for the extraction of nucleic acids using a hybrid paper-plastic device

Nucleic acid-based diagnostics play a crucial role in early and accurate disease detection. However, conventional extraction approaches are expensive, time-consuming, and require complex instrumentation and skilled personnel, which restricts their use in resource-limited settings. To develop and evaluate a simple, low-cost, and equipment-free paper-based microfluidic cassette device for rapid extraction of nucleic acids (DNA and RNA) from biological samples. A paper-plastic cassette was fabricated by laminating filter paper or glass fiber substrates between thermally bonded plastic sheets. The system was tested using Klebsiella pneumoniae (DNA) and HeLa cells (RNA) with different lysis buffer formulations. Extracted nucleic acids were quantified using a Qubit fluorimeter and assessed by gel electrophoresis and PCR amplification. Results were compared with standard commercial extraction kits. Among tested substrates, LF1 combined with lysis buffers 2, 4, and 5 yielded the highest quality DNA and RNA with 260/280 absorbance ratios of approximately 1.8 (DNA) and 2.0 (RNA). The nucleic acid yields were comparable to commercial kits, and the extracted material was suitable for direct downstream applications, including PCR. The developed paper-plastic cassette enables rapid, affordable, and equipment-free extraction of nucleic acids. This platform shows strong potential for point-of-care molecular diagnostics, particularly in low-resource and decentralized healthcare settings.

molecular biology↗