BROADLY REACTIVE APTAMERS TARGETING DIFFERENT GENERA OF COMMON ORGANISMS RELATED TO ACUTE BACTERIAL MENINGITIS
محل انتشار: نوزدهمین کنگره بین المللی میکروب شناسی ایران
سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 386
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شناسه ملی سند علمی:
MEDISM19_018
تاریخ نمایه سازی: 13 مهر 1397
چکیده مقاله:
Background and Aim:Meningitis is one of the ten death leading cause of worldwide infectious diseases. Neisseria meningitides, Haemophilus influenza, and Streptococcus pneumoniae are the most common cause of bacterial meningitis (BM) and finding a new method for detection of BM is an urgent need for clinical treatment.Methods:We applied a new method to isolate single-stranded DNA aptamers that have broad affinity for different bacterial genera. The key point of this method is that targets of interest are changed sequentially at each SELEX cycle. The main plan of SELEX was screening of DNA library for three groups of bacteria containing H. influenza type B, N. meningitides serogroups A, B, C, Y and S. pneumonia serotypes 19A, 6A, 6B. The amplicons were sequentially separated through incubation with each group and combinational cell mixtures as complete SELEX cycle repeated four times. The binding efficiency of the ssDNA pools shows progressive increment estimated by flow cytometry up to 45%. The aptamer pool of round 20th was cloned and transferred in to E.coli Dh5α cells.Results:A total of 93 aptamer transformants were achieved. These aptamers showed the highest affinity and specificity for superior cause of BM and the lowest affinity to other groups of bacteria.Conclusion:These results demonstrate the potential to isolate aptamers with broad affinity to bacterial taxa in different genera and development of diagnostic tools for multiple targets with similar or different structures.
کلیدواژه ها:
DNA aptamer library ، Selection of DNA aptamers ، Affinity binding and recognition ، Cell SELEX for bioanalytical assays ، Detection of bacterial meningitis
نویسندگان
Atefeh Pourdadashi
M.Sc student of microbial biotechnology, Faculty of Basic Science, Department of Biology, Shahed University
Seyed Latif Mousavi Gargari
Professor in Biotechnology , Faculty of Basic Science, Shahed University