Rapid aptasensor capable of simply diagnosing prostate cancer

被引:50
作者
Cha, Timothy [1 ,2 ]
Cho, Sandy [1 ,3 ]
Kim, Young Teck [4 ]
Lee, Ji Hoon [1 ]
机构
[1] Luminescent MD LLC, Hagerstown, MD 21742 USA
[2] Thomas Jefferson High Sch Sci & Technol, Alexandria, VA 22312 USA
[3] McLean High Sch, Mclean, VA 22101 USA
[4] Virginia Polytech Inst & State Univ, Dept Wood Sci & Forest Prod, Blacksburg, VA 24061 USA
关键词
G-rich DNA aptamer; Prostate specific antigen (PSA); Diagnosis; Guanine chemiluminescence; Chemiluminescent resonance energy transfer (CRET); 3,4,5-trimethoxylphenylglyoxal (TMPG); CHEMILUMINESCENCE; APTAMER; PSA; IMMUNOSENSOR;
D O I
10.1016/j.bios.2014.06.015
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Using guanine (G)-rich DNA aptamer-conjugated 6-carboxyfluorescein (6-FAM) capable of rapidly capturing prostate specific antigen (PSA) in human serum, cost-effective and simple biosensor with guanine chemiluminescence detection was developed for early diagnosis of prostate cancer. Free G-rich DNA aptamer-conjugated 6-FAM emits bright light in guanine chemiluminescence reaction based on the principle of chemiluminescent resonance energy transfer (CRET). However, G-rich DNA aptamer-conjugated 6-FAM bound with PSA cannot emit light because PSA acts as a strong interference in CRET between 6-FAM and high-energy intermediate formed from the reaction of 3,4,5-trimethoxylphenylglyoxal (TMPG) and guanine of G-rich DNA aptamer. A chemiluminescent biosensor, developed using the different properties of G-rich DNA aptamer-conjugated 6-FAM in the absence and presence of PSA in guanine chemiluminescence reaction, was able to quantify trace levels of PSA in human serum within 30 min without time-consuming and complicated procedures (e.g., multiple incubation and washings) required for conventional immunoassays operated with expensive and intractable antibodies. The limit of detection of chemiluminescent biosensor having a wide linear dynamic range (1.9-125 ng/ml) was 1.0 ng/ml. The excellent correlation (R=0.985) between chemiluminescent biosensor and conventional enzyme immunoassay indicates that the accurate, precise, and rapid chemiluminescent biosensor can be applied as a new method for early diagnosis of prostate cancer. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:31 / 37
页数:7
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