Direct detection of hyaluronidase in urine using cationic gold nanoparticles: a potential diagnostic test for bladder cancer.
Hyaluronidase (HAase) was reported as a urinary marker of bladder cancer. In this study, a simple colorimetric gold nanoparticle (AuNP) assay was developed for rapid and sensitive detection of urinary HAase activity. Charge interaction between polyanionic hyaluronic acid (HA) and cationic AuNPs stabilized with cetyl trimethyl ammonium bromide (CTAB) led to formation of gold aggregates and a red to blue color shift. HAase digests HA into small fragments preventing the aggregation of cationic AuNPs. The nonspecific aggregation of AuNPs in urine samples was overcome by pre-treatment of samples with the polycationic chitosan that was able to agglomerate all negatively charged interfering moieties before performing the assay. The developed AuNP assay was compared with zymography for qualitative detection of urinary HAase activity in 40 bladder carcinoma patients, 11 benign bladder lesions patients and 15 normal individuals, the assay sensitivity was 82.5% vs. 65% for zymography, while the specificity for both assays was 96.1%. The absorption ratio, A530/A620 of the reacted AuNP solution was used to quantify the HAase activity. The best cut off value was 93.5 μU/ng protein, at which the sensitivity was 90% and the specificity was 80.8%.The developed colorimetric AuNP HAase assay is simple, inexpensive, and can aid noninvasive diagnosis of bladder cancer.
Biochemistry Department, Faculty of Pharmacy, Misr University for Science and Technology (MUST), 6th October City, Egypt.
Ahmed Ibrahim
AI
Nossier
Oncology Diagnostic Unit, Medical Biochemistry & Molecular Biology Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt.
Sanaa
S
Eissa
Biochemistry Department, Faculty of Pharmacy, Cairo University, Giza, Egypt.
Manal Fouad
MF
Ismail
Biochemistry Department, Faculty of Pharmacy, Cairo University, Giza, Egypt.
Mohamed Ahmed
MA
Hamdy
Department of Chemistry & Yousef Jameel Science & Technology Research Center, The American University in Cairo, New Cairo, Egypt. Electronic address: hazzazy@aucegypt.edu.
Hassan Mohamed El-Said
HM
Azzazy
D014408:Biomarkers, Tumor / Q000652:urine*; D003124:Colorimetry / Q000191:economics / Q000379:methods; D006046:Gold / Q000737:chemistry*; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000652:urine*; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000652:urine*
D014408:Biomarkers, Tumor; D006046:Gold; D006821:Hyaluronoglucosaminidase
Bladder Cancer; Chitosan; Gold Nanoparticles; Hyaluronic acid; Hyaluronidase