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Title: Colorimetric detection of PCA3 in urine for prostate cancer diagnosis using thiol-labeled PCR primer and unmodified gold nanoparticles
Authors: Khin Phyu Pyar Htoo
Vichanan Yamkamon
Sakda Yainoy
Thummaruk Suksrichavalit
Wit Viseshsindh
Warawan Eiamphungporn
Faculty of Medicine, Ramathibodi Hospital, Mahidol University
Mahidol University
University of Medical Technology
Keywords: Biochemistry, Genetics and Molecular Biology;Medicine
Issue Date: 1-Jan-2019
Citation: Clinica Chimica Acta. Vol.488, (2019), 40-49
Abstract: © 2018 Elsevier B.V. Background: PCA3, a non-coding RNA, has been approved as a potential urinary biomarker for prostate cancer. However, PCA3 urine tests have some limitations. Therefore, we developed a colorimetric method for PCA3 detection in urine. Methods: The assay was based on interactions between unmodified gold nanoparticles (AuNPs) and thiolated PCR products. Thiolated PCR products were amplified by RT-PCR using a thiol-labeled primer at the 5′ end. Thiolated products of PCA3 bound to the surface of AuNPs and led to the prevention of salt-induced aggregation (red color). In the absence of the PCR products, AuNPs changed their color from red to blue due to the salt-induced aggregation. These changes were detected by the naked eye and spectrophotometer. Results: Our assay was specific for PCA3 in prostate cancer cell lines with a visual detection limit of 31.25 ng/reaction. The absorption ratio 520/640 nm was linear against PCR product concentration (R 2 = 0.9798) in the reaction. This method is promising for discrimination of prostate cancer patients from both healthy controls and benign prostatic hyperplasia patients according to their urinary PCA3 expression levels. Conclusions: This study established a simple, rapid, sensitive and specific assay for PCA3 detection which may be applicable for prostate cancer diagnosis.
ISSN: 18733492
Appears in Collections:Scopus 2019

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