title: Identification of ssDNA aptamers specific to clinical isolates of Streptococcus mutans strains with different cariogenicity.
abstract: Streptococcus mutans, a Gram-positive facultative anaerobic bacterium, is considered to be a major etiological factor for dental caries. In this study, plaques from dental enamel surfaces of caries-active and caries-free individuals were obtained and cultivated for S. mutans isolation. Morphology examination, biochemical characterization, and polymerase chain reaction were performed to identify S. mutans The cariogenicity of S. mutans strains isolated from clinical specimens was evaluated by testing the acidogenicity, aciduricity, extracellular polysaccharide production, and adhesion ability of the bacteria. Finally, subtractive SELEX (systematic evolution of ligands by exponential enrichment) technology targeting whole intact cells was used to screen for ssDNA aptamers specific to the strains with high cariogenicity. After nine rounds of subtractive SELEX, sufficient pool enrichment was achieved as shown by radioactive isotope analysis. The enriched pool was cloned and sequenced randomly, followed by MEME online and RNA structure software analysis of the sequences. Results from the flow cytometry indicated that aptamers H1, H16, H4, L1, L10, and H19 could discriminate highly cariogenic S. mutans strains from poorly cariogenic strains. Among these, Aptamer H19 had the strongest binding capacity with cariogenic S. mutans strains with a dissociation constant of 69.45 ± 38.53 nM. In conclusion, ssDNA aptamers specific to highly cariogenic clinical S. mutans strains were successfully obtained. These ssDNA aptamers might be used for the early diagnosis and treatment of dental caries.
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China Department of Stomatology, Chinese PLA No. 513 Hospital, Lanzhou 732750, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
authors.affiliation: Department of Stomatology, General Hospital of Shenyang Command, Shenyang 110840, China.
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Special Clinic, School of Stomatology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Donghua
authors.initials: D
authors.lastname: Su
authors.affiliation: Xiaopingdao Division, Dalian Sanatorium of Shenyang Military Region, Dalian 116023, China.
authors.forename: Danyang
authors.initials: D
authors.lastname: Hu
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Hou
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Tongnan
authors.initials: T
authors.lastname: Hu
authors.affiliation: Department of Microbiology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Yang
authors.affiliation: Xiaopingdao Division, Dalian Sanatorium of Shenyang Military Region, Dalian 116023, China.
authors.forename: Yanping
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China 66937964@163.com chubingf@aliyun.com Xqsg@sina.com.cn.
authors.forename: Qing
authors.initials: Q
authors.lastname: Xi
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China 66937964@163.com chubingf@aliyun.com Xqsg@sina.com.cn.
authors.forename: Bingfeng
authors.initials: B
authors.lastname: Chu
authors.affiliation: Department of Stomatology, Chinese PLA General Hospital, Beijing 100853, China 66937964@163.com chubingf@aliyun.com Xqsg@sina.com.cn.
authors.forename: Chenglong
authors.initials: C
authors.lastname: Wang
mesh_terms: D000328:Adult; D052157:Aptamers, Nucleotide / Q000235:genetics; D001422:Bacterial Adhesion; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D003731:Dental Caries / Q000382:microbiology*; D003773:Dental Plaque / Q000382:microbiology; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D052156:SELEX Aptamer Technique; D013045:Species Specificity; D013295:Streptococcus mutans / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics; D055815:Young Adult
chemical_list: D052157:Aptamers, Nucleotide; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded
keywords: SELEX; Streptococcus mutans; aptamers; selection