:: Volume 22, Issue 3 (5-2018) ::
ibj 2018, 22(3): 193-201 Back to browse issues page
Development of a DNA Aptamer for Screening Neisseria meningitidis Serogroup B by Cell SELEX
Kimia Mirzakhani *, Seyed Latif Mousavi Gargari, Iraj Rasooli, Samaneh Rasoulinejad
Background: Artificial oligonucleotides like DNA or RNA aptamers can be used as biodiagnostic alternatives for antibodies to detect pathogens. Comparing to antibodies, artificial oligonucleotides are produced easily at lower costs and are more stable. Neisseria meningitidis, the causative agent of meningitis, is responsible for about 1% of infections in an epidemic period. Specific DNA aptamers that bind to N. meningitidis serogroup B were identified by whole-cell Systemic Evolution of Ligands by EXponential Enrichment (SELEX). Methods: The SELEX begins with a library of labeled ssDNA molecules. After six rounds of selection and two rounds of counter-selection, 60 clones were obtained, of which the binding efficiency of 21 aptamers to the aforementioned bacterium was tested by flow cytometry. Results: The aptamers K3 and K4 showed the highest affinity to N. meningitidis serogroup B and no affinity to N. meningitidis serogroups Y, A, and C, or to other meningitis causing bacteria. The dissociation constant (Kd value) for K3 and K4 were calculated as 28.3 ± 8.9 pM and 39.1 ± 8.6 pM, respectively. K3 aptamer with the lowest Kd was chosen as the main aptamer. K3 could detect N. meningitidis in patients’ cerebrospinal fluid (CSF) samples and in CSF from healthy volunteers inoculated with N. meningitidis serogroup B (ATCC 13090) at 200 and 100 CFU ml-1, respectively. Conclusion: The findings suggest the application of the developed aptamer in specific detection of N. meningitidis serogroup B amongst a group of meningitis causing bacteria. 
Keywords: Aptamer, Flow cytometry, Neisseria meningitidis, Serogroup
Full-Text [PDF 942 kb]      
Type of Study: Full Length | Subject: Related Fields
Add your comments about this article
Your username or Email:


XML     Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Mirzakhani K, Mousavi Gargari S L, Rasooli I, Rasoulinejad S. Development of a DNA Aptamer for Screening Neisseria meningitidis Serogroup B by Cell SELEX. ibj. 2018; 22 (3) :193-201
URL: http://ibj.pasteur.ac.ir/article-1-2190-en.html

Volume 22, Issue 3 (5-2018) Back to browse issues page
Persian site map - English site map - Created in 0.17 seconds with 38 queries by YEKTAWEB 3879