Browsing by Author "SANIA MUNIR"

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    Validation of biomarkers for foodborne pathogens
    (UMT Lahore, 2025-09-29) SANIA MUNIR; SALAHA; KIRAN MURTAZA
    The growing need for rapid and accurate detection of microorganisms in food samples has led to the advancement of previously identified biomarkers for food safety testing. However, their reliability requires comprehensive validation. This study aims to validate previously identified biomarkers for detecting microorganisms in various fresh and spoiled fruit samples, including grapes and banana, using molecular techniques, specifically polymerase chain reaction (PCR). We carried three samples: spoiled banana (SB), spoiled grapes (SG), and fresh banana (FB). Under PCR validation, SB showed positive bands for E. coli and Salmonella typhi, SG showed clear amplification for Brucella melitensis and E. coli, while FB produced weak or negative bands consistent with the absence of pathogens. Among these, the spoiled grapes sample expressed the results most accurately under PCR validation, showing strong and reproducible amplification patterns. Our methodology involved first culturing the spoiled food samples to isolate microorganisms, then it was subjected to standard biochemical testing including catalase, oxidase, urease, citrate, indole, MR-VP, and growth on selective media. These tests confirmed the enzymatic and metabolic characteristics of isolates. The results offered the initial information prior to molecular validation through PCR, followed by PCR amplification using available biomarkers. The PCR assays were performed to detect the target microorganisms with high accuracy. After PCR amplification, results were analyzed through gel electrophoresis. We then compared the results obtained from the biomarkers with those obtained from traditional culture methods. The main objective was to evaluate the specificity, sensitivity, and accuracy of biomarkers. The future of this study lies in extending biomarker validation to a wider range of fruits and food matrices to ensure broader applicability. Incorporating advanced molecular methods such as qPCR, multiplex PCR, and next-generation sequencing could enhance sensitivity and allow simultaneous detection of multiple pathogens. Additionally, adapting these validated biomarkers into portable biosensor-based platforms would enable rapid, on-site screening for foodborne pathogens, improving food safety monitoring at the production and distribution levels. Finally, large-scale validation across diverse environments and real market samples will strengthen the reliability and potential commercialization of these biomarkers as routine diagnostic tools.