Preparation And Identification Of Bladder Cancer Aptamer-MNPs Complex And It's Clinical Application In Urine Cytology

Objective:Evaluate the recognition ability of the bladder cancer aptamer-MNPs complex to bladder urothelial cancer cells,which is prepared by biotin-streptavidin method.Further more,we evaluate the value of bladder cancer aptamer-MNPs complex in urine cytology,and it's clinical application value in non-invasive examination of bladder cancer.Methods:1.Evaluate the recognition and identification ability of bladder cancer aptamer aptl to bladder urothelial carcinoma EJ cells by cell flow cytometry and fluorescence microscope,and determine it's Kd value by setting up aptl concentration gradient.2.Prepare bladder cancer aptamer-MNPs complex by biotin-streptavidin method,and evaluate the recognition and identification ability of the complex to EJ cells of bladder urothelial carcinoma by white light and fluorescence microscopy.3.To evaluate the value of bladder cancer aptamer-MNPs complex in urine cytology,we enrolled 106 patients of our hospital due to suspected bladder tumors for study from March 1,2019 to January 1,2020.The enrolled patients were tested by traditional urine cytology,NMP22 assay,and our bladder cancer aptamer-MNPs complex detection.The value of bladder cancer diagnosis was compared in three methods,cystoscope plus tissue biopsy results were used as the criteria.Calculate the sensitivity and specificity of the three methods respectively,and analyze the results of three methods with pathological grade and tumor stagy of the patients by chi-square test.Evaluate the value of the bladder cancer aptamer-MNPs complex in bladder urothelial cancer diagnosis comprehensively.Results:1.The bladder cancer aptamer apt1 can bind with EJ cells specifically,which hardly bind with SV,293T,BGC-803 cells,and it's Kd value of EJ cells is 32..86±16.86nM.2.The bladder cancer aptamer-MNPs complex can recognize and capture EJ cells specifically,and we can see magnetic beads on the surface of EJ cells obviously,and few magnetic beads on SV and 293T cells.3.This study compared the diagnostic value of three methods in bladder urothelial carcinoma.Bladder cancer aptamer-MNPs complex possess high specificity(94.73%)same as urine cytology assay,while the sensitivity(73.56%)is also superior than traditional urine cytology assay(47.12%)and NMP22 assay(62.06%).4.The positive detection ratio of three methods increased with the increase of tumor stage,and the difference was statistically significant(p<0.05).5.The positive detection ratio of traditional urine cytology increased with the increase of pathological grade,and the difference was statistically significant(p=0.006).The positive detection ratio of bladder cancer aptamer-MNPs complex and NMP22 method increased with the increase of pathological grade slightly,but the difference was not statistically significant(p>0.05).Conclusions:The bladder cancer aptamer-MNPs complex prepared in this study is simple,economical,and reliable.It possesses high sensitivity(73.56%)and high specificity(94.73%)in bladder urothelial carcinoma diagnosis.At the same time,the bladder cancer aptamer-MNPs complex method is easy to operate and the results are intuitive and reliable.It has the potential to diagnose bladder urothelial carcinoma.