Signal enhancement in ELISA: Biotin-streptavidin technology against gold nanoparticles

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• 作者：Thangavel Lakshmipriya ; Ph.D^a ; Subash C.B. Gopinath ; Ph.D^b ; Uda Hashim ; Ph.D^b ; Thean-Hock Tang ; Ph.D^a ; ^{tangth@amdi.usm.edu.my" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Biotin ; ELISA ; GNP ; Streptavidin ; Tuberculosis
• 刊名：Journal of Taibah University Medical Sciences
• 出版年：2016
• 出版时间：October 2016
• 年：2016
• 卷：11
• 期：5
• 页码：432-438
• 全文大小：1431 K

文摘

The aim of this study is to compare the use of biotin-streptavidin and gold nanoparticle (GNP) technologies in enzyme linked immunosorbent assay (ELISA) techniques to improve its sensitivity and accuracy.

absSec_2">Methods

abspara0015">We evaluated two ELISA methods to improve the sensitivity and accuracy. Biotin-streptavidin technology was selected to enhance the ELISA limit of detection due to the high binding affinity of biotin-streptavidin. GNP-conjugated biomolecules were selected to improve detection by ELISA. To evaluate these two methods, the early secreted antigenic target-6 (ESAT-6) from Mycobacterium tuberculosis and the anti-ESAT-6 antibody were used.

absSec_3">Results

abspara0020">The detection limit of ESAT-6 was the same with and without GNP due to the saturation of biotin and streptavidin binding. However, higher absorbance was noticed using GNP only.

absSec_4">Conclusion

abspara0025">The proposed modified ELISA can be used to screen different types of common diseases. Additionally, this study showed how several new techniques can improve the detection and accuracy of ELISA.