Selection of DNA aptamers against Mycobacterium tuberculosis Ag85A, and its application in a graphene oxide-based fluorometric assay

被引：23

作者：

Ansari, Najmeh ^{[1
]}

Ghazvini, Kiarash ^{[2
]}

Ramezani, Mohammad ^{[3
]}

Shahdordizadeh, Mahin ^{[4
]}

Yazdian-Robati, Rezvan ^{[4
]}

Abnous, Khalil ^{[5
]}

Taghdisi, Seyed Mohammad ^{[6
]}

机构：

[1] Mashhad Univ Med Sci, Fac Med, Dept Microbiol & Virol, Mashhad 9177899191, Iran

[2] Mashhad Univ Med Sci, Fac Med, Dept Microbiol & Virol, Buali Res Inst,Antimicrobial Resistance Res Ctr, Mashhad 9177899191, Iran

[3] Mashhad Univ Med Sci, Nanotechnol Res Ctr, Mashhad 9177899191, Iran

[4] Mashhad Univ Med Sci, Sch Pharm, Dept Pharmaceut Biotechnol, Mashhad 9177899191, Iran

[5] Mashhad Univ Med Sci, Pharmaceut Res Ctr, Mashhad 9177899191, Iran

[6] Mashhad Univ Med Sci, Targeted Drug Delivery Res Ctr, Mashhad 9177899191, Iran

来源：

MICROCHIMICA ACTA | 2018年 / 185卷 / 01期

关键词：

SELEX; Serum; Fluorescent assay; Limit of detection; Secretory protein; Quenching; ANTIGEN-85; COMPLEX; DIAGNOSIS; PROTEIN; BIOSENSOR; CULTURE; CFP-10; SELEX;

D O I：

10.1007/s00604-017-2550-3

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

The Mycobacterium Ag85 complex is the major secretory protein of M. tuberculosis. It is a potential marker for early diagnosis of tuberculosis (TB). The authors have identified specific aptamers for Ag85A (FbpA) via protein SELEX using magnetic beads. After twelve rounds of selection, two aptamers (Apt8 and Apt22) were chosen from different groups, and their binding constants were determined by flow cytometry. Apt22 (labeled with Atto 647N) binds to FbpA with high affinity (K-d = 63 nM) and specificity. A rapid, sensitive, and low-cost fluorescent assay was designed based on the use of Apt22 and graphene oxide, with a limit of detection of 1.5 nM and an analytical range from 5 to 200 nM of FbpA.

引用

页数：8