尿液多基因联合检测在尿路上皮癌辅助诊断及疗效评估中应用的初步研究OA

Background and purpose:The early diagnosis of urothelial carcinoma(UC)remains challenging because of the limited sensitivity of urinary cytology and the invasive nature of cystoscopy.This study aimed to establish and validate a urine exfoliated cell-based multi-gene panel assay and to evaluate its clinical value in the auxiliary diagnosis and treatment efficacy assessment of UC.Methods:Urine samples were collected from patients who visited Fudan University Shanghai Cancer Center between February 2025 and February 2026.Pathological diagnosis and staging were based on the 8th edition of the American Joint Committee on Cancer(AJCC)Cancer Staging Manual and the World Health Organization Classification of Tumours of the Urinary System and Male Genital Organs.The inclusion criteria were as follows:① age≥18 years;② definitive pathological diagnosis confirmed by cystoscopic biopsy or surgery;③ complete clinical data.The exclusion criteria were:① suspected multiple primary tumors;② urine samples that do not meet the testing requirements,such as insufficient sample volume,improper storage or transportation,sample contamination,insufficient cell pellet after centrifugation,or DNA concentration/quality failing to meet the assay requirements;③ incomplete clinical data,pathological diagnosis,or key clinical information.This study was approved by the ethics committee of Fudan University Shanghai Cancer Center(ethics approval number:050432-4-2108*),and all patients signed informed consent forms.Multiplex polymerase chain reaction(PCR)was performed to detect FGFR3 and TERT gene mutations and the methylation status of HIST1H4F,NRN1 and POU4F2.Interference testing and precision validation were also conducted.Receiver operating characteristic(ROC)curve analysis was used to evaluate diagnostic performance,and the associations of the multi-gene panel assay with clinicopathological features and treatment efficacy were analyzed.Results:The interference testing and precision validation of the multi-gene panel assay met the predefined requirements.Finally,125 samples were included in the analysis,with an inclusion rate of 75.3%.Among them,55 newly diagnosed and untreated UC patients constituted the case group(including 51 cases of bladder cancer,3 cases of renal pelvic carcinoma and 1 cases of ureteral carcinoma);32 non-UC patients served as the control group(including 19 cases of prostate cancer,10 cases of kidney cancer,1 case of testicular cancer,1 case of endometrial cancer,and 1 case of breast cancer).Additionally,38 UC patients who achieved effective treatment based on clinical evaluation were included for treatment efficacy assessment.The area under the ROC curve(AUC)was 0.926(95%CI:0.850-0.971),with a sensitivity of 87.3%(95%CI:75.5%-94.7%),specificity of 93.8%(95%CI:79.2%-99.2%),positive predictive value of 96.0%(95%CI:85.1%-99.3%),and negative predictive value of 81.1%(95%CI:64.3%-91.4%).No statistically significant difference was observed between multi-gene combined detection results obtained from 10 mL and 30 mL urine samples(P=0.182).The multi-gene panel results were significantly associated with the pathological grade of UC(P<0.001),but not with human epidermal growth factor receptor 2(HER2)expression or the Ki-67 proliferation index(P=0.453 and 0.769,respectively).The positive rate was 87.3%(48/55)among newly diagnosed UC patients and decreased to 10.5%(4/38)among patients who achieved effective treatment,with a statistically significant difference between the two groups(P<0.05).Conclusion:The urine exfoliated cell-based multi-gene panel assay showed high diagnostic sensitivity and specificity and may have potential clinical value in the auxiliary diagnosis,pathological grade indication,and treatment efficacy monitoring of UC.A urine sample volume of only 10 mL was sufficient for testing,which may facilitate its clinical application and promotion.