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

Content Provider	Scilit
Author	Kong, Xiao-Li Qi, Hui Zhou, Han-Xin Ren, Li-Li Deng, Chong-Yan Li, Fu-Rong
Copyright Year	2010
Abstract	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. 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). Each IgG-Au-DNA could be covered with 138+/-47 oligonucleotides and 11+/-3 antibodies. The IgG-M could bind 118 mug 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). 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.
Related Links	https://edoc.hu-berlin.de/bitstream/18452/10248/1/cclm.2010.041.pdf
ISSN	14346621
e-ISSN	14374331
DOI	10.1515/cclm.2010.041
Journal	Clinical Chemistry and Laboratory Medicine (CCLM)
Issue Number	2
Volume Number	48
Language	English
Publisher	Walter de Gruyter GmbH
Publisher Date	2010-01-01
Access Restriction	Open
Subject Keyword	Clinical Chemistry and Laboratory Medicine (cclm) Medical Laboratory Technology Enzyme Linked Immunosorbent Assay Nucleic Acid Barcode Prostate-specific Antigen Radioimmunoassay Sensitive Immunoassay Journal: Clinical Chemistry and Laboratory Medicine (CCLM), Vol- 48
Content Type	Text
Resource Type	Article
Subject	Biochemistry (medical) Clinical Biochemistry