Microplate magnetic chemiluminescence immunoassay for detecting urinary survivin in bladder cancer

被引:20
作者
Chang, Yanli [1 ]
Xu, Jianjun [1 ]
Zhang, Qingyun [1 ]
机构
[1] Peking Univ, Dept Clin Lab, Key Lab Carcinogenesis & Translat Res, Minist Educ,Canc Hosp & Inst, 52 Fucheng Rd, Beijing 100142, Peoples R China
关键词
microplate magnetic chemiluminescence immunoassay; survivin; bladder cancer; tumor markers; ENZYME-IMMUNOASSAY; RAPID DETECTION; CELL-CARCINOMA; MESSENGER-RNA; HUMAN SERUM; EXPRESSION; BIOMARKERS; RECURRENCE; PROGNOSIS; DIAGNOSIS;
D O I
10.3892/ol.2017.6675
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Survivin is a tumor marker for bladder cancer; however the role of urinary survivin levels has not been fully elucidated due to the limitations of current detection methods. Based on two survivin-specific monoclonal antibodies (McAbs) already confirmed through enzyme linked immunosorbent assays, the present study aimed to establish a microplate magnetic chemiluminescence immunoassay (CLIA) for the detection of urinary survivin levels and evaluate its application for the diagnosis of patients with bladder cancer. Horseradish peroxidase and biotin conjugates were used to label two different anti-survivin McAbs, respectively. The labeled antibodies combined with survivin to form a sandwiched immune complex. The streptavidin magnetic particles (MPs) served as the solid phase and the separator. The relevant parameters involved in the immunoassay, including the immunoassay reagents used and the physicochemical parameters were optimized. Then, urine samples from 130 patients with bladder cancer and 113 healthy controls were detected, and analyzed using the established method. The method was linear to 1,000 ng/ml survivin with a detection limit of 0.83 ng/ml. The intra-and inter-assay coefficients of variation were <8, and <11%, respectively. The concentration of diluted survivin and the dilution ratios gave a linear correlation of 0.9989. The results demonstrated that the urinary survivin levels in patients with bladder cancer were significantly higher (P<0.001) compared with that in healthy controls. At a survivin concentration of 2.0884 ng/ml, the sensitivity and specificity were 86.9 and 61.9%, respectively. Furthermore, the urinary survivin levels were positively correlated with metastatic stage, histological stage and recurrence (P<0.01). In conclusion, the present study preliminarily proposed a microplate magnetic CLIA for survivin detection and further evaluated the value of urinary survivin as a diagnostic marker for bladder cancer.
引用
收藏
页码:4043 / 4052
页数:10
相关论文
共 48 条
[1]  
Abd El-Hakim TF, 2014, ANAL QUANT CYTOL, V36, P121
[2]   The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins [J].
Adachi, Jun ;
Kumar, Chanchal ;
Zhang, Yanling ;
Olsen, Jesper V. ;
Mann, Matthias .
GENOME BIOLOGY, 2006, 7 (09)
[3]   Emmprin and survivin predict response and survival following cisplatin-containing chemotherapy in patients with advanced bladder cancer [J].
Als, Anne B. ;
Dyrskjot, Lars ;
von der Maase, Hans ;
Koed, Karen ;
Mansilla, Francisco ;
Toldbod, Helle E. ;
Jensen, Jens L. ;
Ulhoi, Benedicte P. ;
Sengelov, Lisa ;
Jensen, Klaus M. E. ;
Orntoft, Torben F. .
CLINICAL CANCER RESEARCH, 2007, 13 (15) :4407-4414
[4]  
Altieri Dario C, 2003, Prog Cell Cycle Res, V5, P447
[5]   EAU Guidelines on Non-Muscle-invasive Urothelial Carcinoma of the Bladder: Update 2016 [J].
Babjuk, Marko ;
Boehle, Andreas ;
Burger, Maximilian ;
Capoun, Otakar ;
Cohen, Daniel ;
Comperat, Eva M. ;
Hernandez, Virginia ;
Kaasinen, Eero ;
Palou, Joan ;
Roupret, Morgan ;
van Rhijn, Bas W. G. ;
Shariat, Shahrokh F. ;
Soukup, Viktor ;
Sylvester, Richard J. ;
Zigeuner, Richard .
EUROPEAN UROLOGY, 2017, 71 (03) :447-461
[6]  
Budman LI, 2008, CUAJ-CAN UROL ASSOC, V2, P212
[7]  
Chen D, ONCOLOGY RE IN PRESS
[8]   IMMUNOMAGNETIC SEPARATION OF SALMONELLA FROM FOODS AND THEIR DETECTION USING IMMUNOMAGNETIC PARTICLE (IMP)-ELISA [J].
CUDJOE, KS ;
HAGTVEDT, T ;
DAINTY, R .
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, 1995, 27 (01) :11-25
[9]   Protein microarray for the analysis of human melanoma biomarkers [J].
Domnanich, Patrick ;
Sauer, Ursula ;
Pultar, Johanna ;
Preininger, Claudia .
SENSORS AND ACTUATORS B-CHEMICAL, 2009, 139 (01) :2-8
[10]   Diagnostic Evaluation of Apoptosis Inhibitory Gene and Tissue Inhibitor Matrix Metalloproteinase-2 in Patients with Bladder Cancer [J].
Eissa, Sanaa ;
Shabayek, Marwa I. ;
Ismail, Manal F. ;
El-Allawy, Raouf M. ;
Hamdy, Mohamed A. .
IUBMB LIFE, 2010, 62 (05) :394-399