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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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1437-4331
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Volume 48, Issue 2 (Feb 2010)

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A novel sensitive immunoassay by nucleic acid barcode dot and its application in the detection of prostate-specific antigen

Xiao-Li Kong
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
/ Hui Qi
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
/ Han-Xin Zhou
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
/ Li-Li Ren
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
/ Chong-Yan Deng
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
/ Fu-Rong Li
  • Clinical Medical Research Center, The 2nd Clinic Medicine College (Shenzhen People's Hospital), Jinan University, Shenzhen, P.R. China
Published Online: 2009-12-10 | DOI: https://doi.org/10.1515/CCLM.2010.041

Abstract

Background: The sensitivity and selectivity of traditional methods limits ultramicro detection of proteins. Bio-barcode amplification detection methods based on nanotechnology enables ultramicro detection of protein. However, bio-barcode amplification detection depends on the oligonucleotides being fixed on a glass chip. It also requires specialized equipment, which limits its application. We introduce a nano-nucleic acid barcode dot detection technology to determine ultramicro concentrations of protein. The method is simple, quick and accurate.

Methods: Magnetic probe (IgG-M) and dual-labeled gold nanoparticle bio-probe (IgG-Au-DNA) were prepared. Protein was captured using a sandwich assay technique and magnetic separation was used. The DNA barcode was released with dithiothreitol (DTT) and detected directly without the requirement for polymerase chain reaction (PCR). Serum prostate-specific antigen (PSA) from 135 patients was detected with this method and compared with enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA).

Results: Each IgG-Au-DNA could be covered with 138±47 oligonucleotides and 11±3 antibodies. The IgG-M could bind 118 μg of antibody per mg. The sensitivity of nano-nucleic acid barcode dot detection technology might allow detection of 1 fg/mL. There were no significant differences in serum PSA from 135 patients when comparing the three methods (compared with ELISA, r=0.950; and with RIA, r=0.967).

Conclusions: The nucleic acid barcode dot method does not require special equipment or complex procedures, but its detection limit is 2–3 orders of magnitude lower than ELISA.

Clin Chem Lab Med 2010;48:279–83.

Keywords: enzyme linked immunosorbent assay; nucleic acid barcode; prostate-specific antigen; radioimmunoassay; sensitive immunoassay

About the article

Corresponding author: Fu-Rong Li, Clinical Medical Research Center, The 2nd Clinic Medicine College, Jinan University, No. 1017 Dongmen North Road, Shenzhen 518020, Guangdong Province, P.R. China Phone: +86-755-25533018, Fax: +86-755-25533497,


Received: 2009-08-17

Accepted: 2009-10-06

Published Online: 2009-12-10

Published in Print: 2010-02-01


Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2010.041. Export Citation

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