DNA aptamers for the detection of Haemophilus influenzae type b by cell SELEX

European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology
F S BitarafS L Mousavi Gargari

Abstract

Haemophilus influenzae type b (Hib) causes acute bacterial meningitis (ABM) in children, with a mortality rate of about 3-6 % of the affected patients. ABM can lead to death during a period of hours to several days and, hence, rapid and early detection of the infection is crucial. Aptamers, the short single-stranded DNA or RNA with high affinity to target molecules, are selected by a high-flux screening technique known as in vitro screening and systematic evolution of ligands by exponential enrichment technology (SELEX). In this study, whole-cell SELEX was applied for the selection of target-specific aptamers with high affinity to Hib. ssDNA aptamers prepared by lambda exonuclease were incubated with the target cells (Hib). The aptameric binding rate to Hib was characterized for binding affinity after seven SELEX rounds by flow cytometry. The aptamers with higher binding affinity were cloned. Four of 68 aptamer clones were selected for sequencing. The dissociation constant (Kd) of the high-affinity aptamer clones 45 and 63 were 47.10 and 28.46 pM, respectively. These aptamers did not bind to other bacterial species, including the seven meningitis-causing bacteria. They showed distinct affinity to various H. influenzae strains o...Continue Reading

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Citations

May 29, 2016·Journal of Biotechnology·Samaneh Rasoulinejad, Seyed Latif Mousavi Gargari
Oct 12, 2016·Biosensors & Bioelectronics·Paria PashazadehMiguel de la Guardia
Jan 26, 2017·Canadian Journal of Microbiology·Hanif AlfavianArvin Medhat
Jun 27, 2020·Applied Microbiology and Biotechnology·Fernanda Raquel Wust SchmitzDachamir Hotza
Jan 29, 2021·Analytical and Bioanalytical Chemistry·Lin ZhuChaoyong Yang

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