Monitoring Of Bladder Cancer Research Articles

Genetic and epigenetic characteristics of non-muscle invasive and muscle invasive bladder cancer in patients infected by human papillomavirus: literature review

Infectious diseases and chronic inflammation are important risk factors for the development of malignant tumors in humans. One of the key infectious agents involved in human oncogenesis is the human papillomavirus (HPV). Non-muscle invasive bladder cancer is defined as a superficial neoplasia limited to the mucosa, aggravated by recurrence in 80 % of cases and progression in 30 % of cases. The development of this disease is associated with the influence of various carcinogenic agents, including HPV. Currently, a direct relationship has been revealed between the presence of viral DNA in the tumor tissue of the bladder and markers of proliferative activity, angiogenesis, and apoptosis factors. More and more researchers believe in the involvement of the virus in the development of recurrent forms of bladder cancer and the emergence of its invasive/poorly differentiated forms. Improving the diagnosis and postoperative monitoring of non-muscle invasive and muscle invasive bladder cancer is not possible without the improvement of minimally invasive molecular methods, which requires an understanding of the molecular mechanisms of HPV-associated carcinogenesis.Therefore, this review focuses on the analysis of the molecular mechanisms of HPV effect on progression of non-muscle invasive and muscle invasive bladder cancer. The features of miRNA expression in patients with papillomavirus infection of high oncogenic risk types and non-muscle invasive or muscle invasive bladder cancer are considered in detail. In particular, the role of miR-34а, -218, -20a, -424, -200a, -205-5p, -944, -100, -99a, -202, -30a, -145-5p, -195 and -199a-5 is described in the development and progression of bladder cancer. The mechanisms of disruption in the functioning of key cell signaling pathways during HPV integration in patients with bladder cancer, including changes in gene copy number and methylation level, are also considered.However, the number of HPV-positive tumor specimens that have been comprehensively analyzed using genome-wide studies in the literature remains small. Larger patient cohorts would be useful to further refine HPV-associated integration events and genomic changes, as well as to study clinical manifestations of the consequences of these alterations. Further research on the clinical implications of the observed genomic changes is needed to accurately stratify patients for targeted therapy, radiation and chemotherapy.

MiRNAs role in bladder cancer pathogenesis and targeted therapy: Signaling pathways interplay - A review.

Bladder cancer (BC) is the 11th most popular cancer in females and 4th in males. A lot of efforts have been exerted to improve BC patients' care. Besides, new approaches have been developed to enhance the efficiency of BC diagnosis, prognosis, therapeutics, and monitoring. MicroRNAs (miRNAs, miRs) are small chain nucleic acids that can regulate wide networks of cellular events. They can inhibit or degrade their target protein-encoding genes. The miRNAs are either downregulated or upregulated in BC due to epigenetic alterations or biogenesis machinery abnormalities. In BC, dysregulation of miRNAs is associated with cell cycle arrest, apoptosis, proliferation, metastasis, treatment resistance, and other activities. A variety of miRNAs have been related to tumor kind, stage, or patient survival. Besides, although new approaches for using miRNAs in the diagnosis, prognosis, and treatment of BC have been developed, it still needs further investigations. In the next words, we illustrate the recent advances in the role of miRNAs in BC aspects. They include the role of miRNAs in BC pathogenesis and therapy. Besides, the clinical applications of miRNAs in BC diagnosis, prognosis, and treatment are also discussed.

What is the role of CellDetect® in detecting and monitoring bladder cancer?

Cystoscopy is the gold standard method for diagnosing and monitoring bladder cancer but it is costly, invasive, and operator-dependant. The aim is to compare the diagnostic efficacy of CellDetect® with urine cytology based on cystoscopic findings. A total of 181 patients undergoing cystoscopy for bladder cancer follow-up or any reason were studied with cytology and CellDetect® by taking an urine sample before cystoscopy. Patients who had any kind of bladder procedure in less than 1 month, doubtful cystoscopy results, previous pathology of Tis or carcinoma in situ (CIS), urinary stones, and patients with urinary catheters or bladder diversions were excluded. Cytologic and CellDetect® results were compared based on cystoscopic findings and sensitivity and specifity analyses were done for each method. For low-grade tumors, the sensitivity of CellDetect® was 66.7% and the sensitivity of cytology was 16.7% with a significant difference (p < 0.05). For high-grade tumors, there were no significant difference between CellDetect® and cytology. Generally, CellDetect® had better sensitivity in both case and control groups. The promising results of CellDetect® particularly in low-grade tumors gives the potential for this novel stain to go widespread. Larger series, meta-analyses, and reviews need to support this topic in order to put CellDetect® into daily practice.

UroVysionTM Fluorescence In Situ Hybridization in Urological Cancers: A Narrative Review and Future Perspectives.

UroVysionTM is a fluorescence in situ hybridization assay that was developed for the detection of bladder cancer (UC accounted for 90%) in urine specimens. It consists of fluorescently labeled DNA probes to the pericentromeric regions of chromosomes 3, 7, 17 and to the 9p21 band location of the P16 tumor suppressor gene, which was approved by the Food and Drug Administration (FDA) in 2001 and 2005, respectively, for urine detection in patients with suspected bladder cancer and postoperative recurrence monitoring. Furthermore, recent studies also demonstrated that U-FISH was useful for assessing superficial bladder cancer patients' response to Bacillus Calmette-Guérin therapy and in detecting upper tract urothelial carcinoma. Therefore, positive U-FISH was well known to urologists as a molecular cytogenetic technique for the detection of UC. However, with the continuous enrichment of clinical studies at home and abroad, U-FISH has shown a broader application space in the detection of various urinary primary tumors and even metastatic tumors. This review focuses on summarizing the research status of U-FISH in UC, non-urothelial carcinoma and metastatic tumor, so as to strengthen urologists' more comprehensive understanding of the application value of U-FISH and better complete the accurate diagnosis and treatment of urological cancers.

Multicentric validation of diagnostic tests based on BC-116 and BC-106 urine peptide biomarkers for bladder cancer in two prospective cohorts of patients.

Non-invasive urine-based biomarkers can potentially improve current diagnostic and monitoring protocols for bladder cancer (BC). Here we assess the performance of earlier published biomarker panels for BC detection (BC-116) and monitoring of recurrence (BC-106) in combination with cytology, in two prospectively collected patient cohorts. Of the 602 patients screened for BC, 551 were found eligible. For the primary setting, 73 patients diagnosed with primary BC (n = 27) and benign urological disorders, including patients with macroscopic haematuria, cystitis and/or nephrolithiasis (n = 46) were included. In total, 478 patients under surveillance were additionally considered (83 BC recurrences; 395 negative for recurrence). Urine samples were analysed with capillary electrophoresis-mass spectrometry. The biomarker score was estimated via support vector machine-based software. Validation of BC-116 biomarker panel resulted in 89% sensitivity and 67% specificity (AUCBC-116 = 0.82). A diagnostic score based on cytology and BC-116 resulted in good (AUCNom116 = 0.85) but not significantly better performance (P = 0.5672). A diagnostic score including BC-106 and cytology was evaluated (AUCNom106 = 0.82), significantly outperforming both cytology (AUCcyt = 0.72; P = 0.0022) and BC-106 (AUCBC-106 = 0.67; P = 0.0012). BC-116 biomarker panel is a useful test for detecting primary BC. BC-106 classifier integrated with cytology showing >95% negative predictive value, might be useful for decreasing the number of cystoscopies during surveillance.

P17 - Multicentric validation of nomograms based on urine peptide biomarkers for bladder cancer diagnostics and monitoring in two prospective cohorts of patients

P17 - Multicentric validation of nomograms based on urine peptide biomarkers for bladder cancer diagnostics and monitoring in two prospective cohorts of patients

Discovery and validation of bladder cancer related excreted nucleosides biomarkers by dilution approach in cell culture supernatant and urine using UHPLC-MS/MS

The exploration of nucleoside changes in human biofluids has profound potential for cancer diagnosis. Herein, we developed a rapid methodology to quantify 17 nucleosides by UHPLC-MS/MS. Five pairs of isomers were successfully separated within 8 min. The ME was mostly eliminated by sample dilution folds of 1000 for urine and 40 for CCS. The optimized method was firstly applied to screen potential nucleoside biomarkers in CCS by comprising bladder cancer cell lines (5637 and T24) and normal human bladder cell line SV-HUC-1 together with student's t-test and OPLS-DA. Nucleosides with significant differences in the supernatant of urine samples were also uncovered comparing BCa with the non-tumor group, as well as a comparison of BCa recurrence group with the non-recurrence group. By intersecting the differential nucleosides in CCS and urine supernatant, and then further confirmed using validation sets, the combination of m3C and m1A with AUC of 0.775 was considered as a potential biomarker for bladder cancer diagnosis. A panel of m3C, m1A, m1G, and m22G was defined as potential biomarkers for bladder cancer prognosis with an AUC of 0.819. Above all, this method provided a new perspective for diagnosis and recurrence monitoring of bladder cancer. SignificanceThe exploration of nucleoside changes in body fluids has profound potential for the diagnosis and elucidation of the pathogenesis of cancer. In this study, we developed a rapid methodology for the simultaneous quantitative determination of 17 nucleosides in the supernatant of cells and urine samples using UHPLC-MS/MS to discover and validate bladder cancer related excreted nucleoside biomarkers. The results of this paper provide a new strategy for diagnosis and postoperative recurrence monitoring of bladder cancer and provide theoretical support for the exploration of its pathogenesis.

Immune status for monitoring and treatment of bladder cancer.

The high recurrence rate of non-muscle invasive bladder cancer (BC) and poor prognosis of advanced BC are therapeutic challenges that need to be solved. Bacillus Calmette-Guerin (BCG) perfusion was the pioneer immunotherapy for early BC, and the discovery of immune checkpoint inhibitors has created a new chapter in the treatment of advanced BC. The benefit of immunotherapy is highly anticipated, but its effectiveness still needs to be improved. In this review, we collated and analysed the currently available information and explored the mechaisms by which the internal immune imbalance of BC leads to tumour progression. The relationship between immunity and progression and the prognosis of BC has been explored through tests using body fluids such as blood and urine. These analytical tests have attempted to identify specific immuyne cells and cytokines to predict treatment outcomes and recurrence. The diversity and proportion of immune and matrix cells in BC determine the heterogeneity and immune status of tumours. The role and classification of immune cells have also been redefined, e.g., CD4 cells having recognised cytotoxicity in BC. Type 2 immunity, including that mediated by M2 macrophages, Th2 cells, and interleukin (IL)-13, plays an important role in the recurrence and progression of BC. Pathological fibrosis, activated by type 2 immunity and cancer cells, enhances the rate of cancer progression and irreversibility. Elucidating the immune status of BC and clarifying the mechanisms of action of different cells in the tumour microenvironment is the research direction to be explored in the future.

Detecting and monitoring bladder cancer with exfoliated cells in urine.

Current methods for the diagnosis and monitoring of bladder cancer are invasive and have suboptimal sensitivity. Liquid biopsy as a non-invasive approach has been capturing attentions recently. To explore the ability of urine-based liquid biopsy in detecting and monitoring genitourinary tumors, we developed a method based on promoter-targeted DNA methylation of urine sediment DNA. We used samples from a primary bladder cancer cohort (n=40) and a healthy cohort (n=40) to train a model and obtained an integrated area under the curve (AUC) > 0.96 in the 10-fold cross-validation, which demonstrated the ability of our method for detecting bladder cancer from the healthy. We next validated the model with samples from a recurrent cohort (n=21) and a non-recurrent cohort (n=19) and obtained an AUC > 0.91, which demonstrated the ability of our model in monitoring the progress of bladder cancer. Moreover, 80% (4/5) of samples from patients with benign urothelial diseases had been considered to be healthy sample rather than cancer sample, preliminarily demonstrating the potential of distinguishing benign urothelial diseases from cancer. Further analysis basing on multiple-time point sampling revealed that the cancer signal in 80% (4/5) patients had decreased as expected when they achieved the recurrent-free state. All the results suggested that our method is a promising approach for noninvasive detection and prognostic monitoring of bladder cancer.

106P Comparative analysis of urinary and tissue tumor DNA in muscle-invasive bladder cancer by boosted whole-exome sequencing

Urinary tumor DNA profiling is promising for the diagnosis, monitoring, and treatment stratification of bladder cancer. However, previous studies are mainly using a targeted panel next-generation sequencing (NGS) approach, which is limited to predefined genes and thus lacks comprehensiveness. Here, we apply the boosted whole-exome sequencing (WES) to urinary and tissue tumor DNA in muscle-invasive bladder cancer (MIBC) to comprehensively compare the mutation profiles in matched urine and tissue samples.

Biomarkers for bladder cancer diagnosis, prognosis, monitoring and treatment.

Biomarkers for bladder cancer diagnosis, prognosis, monitoring and treatment.

Chemical tools to monitor bladder cancer progression

Background Bladder cancer (BCa) is the most common cancer of the urinary system. Due to its high incidence and recurrence, as well as limited progress in the effective treatment, BCa is a challenge for today's medicine. Materials and Methods We used a set of chromogenic substrates to differentiate between the stages of bladder cancer progression (G1 (n = 10), G2 (n = 10), G3 (n = 10)). The proteolytic activity in individual the urine samples was determined by absorbance measurements. Then inhibitors of particular classes of enzymes were used to determine which enzymes dominate at a given stage of the neoplastic disease. Results The specific activity of enzymes in the urine of patients with confirmed bladder cancer was determined separately for three (G1, G2, G3) stages of the disease development. What is more, no activity was observed in urine of healthy people (n = 10). Discussion Research shows that specific enzymes are associated with the development of specific stages of cancer. We suspect that the differences in the proteolytic activity of urine samples are due to the presence of a different set of enzymes that are directly related to the particular stage of the disease. Conclusion We obtained three substrates for monitoring individual stages of bladder cancer development.

Urinary Markers for Bladder Cancer Diagnosis and Monitoring.

Hematuria is a typical symptom of bladder cancer which enables early detection of bladder cancer. However, reliable diagnostic tools for bladder cancer using urine samples or other non-invasive methods are lacking. Tremendous attempts have been tried and revealed fancy works to convey definitive diagnostic power using urine samples. In this paper, we reviewed urinary markers for bladder cancer and compared their efficacies.

Highly Sensitive and Specific Detection of Bladder Cancer via Targeted Ultra-deep Sequencing of Urinary DNA

BackgroundThere is an unmet need for an accurate, validated, noninvasive test for diagnosing and monitoring bladder cancer (BC). Detection of BC-associated mutations in urinary DNA via targeted deep sequencing could meet this need. ObjectiveTo test the ability of mutational analysis of urinary DNA to noninvasively detect BC within the context of haematuria investigations and non–muscle-invasive BC (NMIBC) surveillance. Design, setting, and participantsCapture-based ultra-deep sequencing was performed for 443 somatic mutations in 23 genes in 591 urine cell-pellet DNAs from haematuria clinic patients and 293 from NMIBC surveillance patients. Variant calling was optimised to minimise false positives using urine samples from 162 haematuria clinic patients without BC. Outcome measurements and statistical analysisThe sensitivity and specificity for BC diagnosis were determined. Results and limitationsMutational analysis of urinary DNA detected 144 of the 165 haematuria patients diagnosed with incident BC from two independent cohorts, yielding overall sensitivity of 87.3% (95% confidence interval [CI] 81.2–92.0%) at specificity of 84.8% (95% CI 79.9–89.0%). The sensitivity was 97.4% for grade 3, 86.5% for grade 2, and 70.8% for grade 1 BC. Among NMIBC surveillance patients, 25 out of 29 recurrent BCs were detected, yielding sensitivity of 86.2% (95% CI 70.8–97.7%) at specificity of 62.5% (95% CI 56.1–68.0%); a positive urine mutation test in the absence of clinically detectable disease was associated with a 2.6-fold increase in the risk of future recurrence. The low number of recurrences in the NMIBC surveillance cohort and the lower sensitivity for detecting grade 1 pTa BC are limitations. ConclusionsDetection of mutations in a small panel of BC-associated genes could facilitate noninvasive BC testing and expedite haematuria investigations. Following further validation, the test could also play a role in NMIBC surveillance. Patient summaryIdentification of alterations in genes that are frequently mutated in bladder cancer appears to be a promising strategy for detecting disease from urine samples and reducing reliance on examination of the bladder via a telescopic camera inserted through the urethra.

Xpert bladder cancer monitor in surveillance of bladder cancer: Systematic review and meta-analysis.

There is a need for sensitive and specific biomarkers for detecting recurrences in patients with non-muscle invasive bladder cancer (NMIBC) on surveillance. Xpert bladder cancer (BC) monitor is the latest rapid in vitro qualitative test that detects expression of 5 mRNAs using a GeneXpert instrument. The primary aim of this review was to systematically review and pool the data regarding the diagnostic performance of Xpert BC in patients with NMIBC. Systematic literature search using 4 electronic databases (PubMed, EMbase, Scopus and Web of science) was performed. Pooled sensitivity, specificity and diagnostic odds ratio (DOR) were estimated using DerSimonian-Laird random-effects model. Standard Preferred Reporting Items for Systematic reviews and Meta-analysis guidelines were followed and the study protocol was registered with PROSPERO (CRD42021249762). In this review, 11 prospective studies with 2,896 patients were included. Pooled sensitivity and specificity for Xpert BC were 73% (95% confidence interval (CI) 65%-80%) and 77% (95% CI 69%-84%) respectively. Area under curve (AUC) for Xpert BC monitor was 0.81 (0.78-0.84) respectively. Subgroup analysis from 7 studies for patients with high-grade recurrence revealed sensitivity, specificity and AUC of 0.86 (0.77-0.92), 0.78 (0.75-0.81) and 0.87 (0.84-0.90) respectively. Similar analysis for patients with low-grade recurrence revealed sensitivity, specificity and AUC of 0.58 (0.47-0.68), 0.79 (0.75-0.82) and 0.79 (0.75-0.82) respectively. Xpert BC monitor has overall acceptable diagnostic accuracy. Sensitivity is higher for high-grade disease for detecting recurrences in patients with NMIBC on surveillance.

Xpert Bladder Cancer Monitor May Avoid Cystoscopies in Patients Under "Active Surveillance" for Recurrent Bladder Cancer (BIAS Project): Longitudinal Cohort Study.

ObjectivesTo test the hypothesis that patients under active surveillance (AS) for Non-muscle Invasive Bladder Cancer (NMIBC) who were negative on longitudinal re-testing by the Xpert® Bladder Cancer Monitor (Xpert BC Monitor) assay may avoid unnecessary cystoscopies and urine cytology (UC).Subjects/Patients (or Materials) and MethodsThis is a prospective cohort study of patients enrolled in the AS protocol for recurrent NMIBC (Bladder Cancer Italian Active Surveillance, BIAS project), whose urine samples were analyzed by Xpert BC Monitor upon entry in the study (T0). Patients who had a negative Xpert test and did not fail AS, underwent additional Xpert tests after 4 (T1), 8 (T2), and 12 (T3) months. The clinical utility of Xpert was assessed by determining the number of cystoscopies and UC that could be avoided within 1 year.ResultsOverall, 139 patients were tested with Xpert at T0. Median follow-up was 23 (IQR 17–27) months. Sixty-eight (48.9%) patients failed AS, 65 (46.7%) are currently on AS, and 6 (4.3%) were lost at follow-up. At T0 57 (41.0%) patients had a negative test and 36 (63.2%) are still in AS. In patients with 2 consecutives negative Xpert tests, we could have avoided 73.9% of unnecessary cystoscopies, missing 26.4% failure, up to avoid all cystoscopies with 4 negative tests missing only 12% of failure. All the patients with negative Xpert had negative UC. Failure-free-survival at median follow-up (23 month) stratified for having 0, 1, or ≥2 negative tests was 67.0, 55.1. and 84.1, respectively.ConclusionOur findings suggest that Xpert BC Monitor assay, when it is longitudinally repeated, could significantly reduce the number of unnecessary cystoscopies and UC during their follow-up.

The Role of New Technologies in the Diagnosis and Surveillance of Non-Muscle Invasive Bladder Carcinoma: A Prospective, Double-Blinded, Monocentric Study of the XPERT© Bladder Cancer Monitor and Narrow Band Imaging© Cystoscopy

Simple SummaryPatients with bladder cancer (BC) require close follow-up with white-light cystoscopy (WLC) and cytology. In this study, we sought to assess (a) the performance of a novel cystoscopy technology based on Narrow Band Imaging© (NBI), and (b) a new urine test (XPERT© Bladder Cancer Monitor, XBCM) that detects cancer proteins. We compared these to the established standard follow-up diagnostics. Our study showed that NBI cystoscopy does not provide any additional benefit over standard WLC. However, the XBCM urine test performed particularly well in instances of aggressive high-grade tumor recurrence. Therefore, XBCM may have enhanced utility in the early detection of potentially harmful BC recurrence.Follow-up is essential for the early detection of recurrent non-muscle invasive bladder cancers (NMIBC). This study investigates the clinical relevance of new diagnostic tools such as an mRNA-based urine test (XPERT© Bladder Cancer Monitor, XBCM) and Narrow Band Imaging© (NBI) and compares them with the established follow-up diagnostics (white-light cystoscopy (WLC) and urine cytology). This was a prospective, double-blind, single-center study that involved patients undergoing NMIBC screening at a tertiary care center. Enrollment occurred between January 2018 and March 2020. In addition to standard care (WLC, cytology, and ultrasound), patients underwent XBCM urine testing and NBI cystoscopy. In total, 301 WLCs were performed; through this, 49 patients demonstrated NMIBC recurrence. NBI cystoscopy was congruent with WLC in all patients. Cytology showed a sensitivity (SE) and specificity (SP) of 27% and 97% (PPV: 65%; NPV 87%), respectively, whereas XBCM showed SE and SP of 58% and 89%, respectively (PPV: 51%; NPV: 92%; AUC: 0.79 (0.716–0.871)). Subgroup analysis showed improved SE and similar SP (PPV, NPV) for high grade (HG) recurrence, with a SE of 74% and SP of 89% (39%, 97%). NBI cystoscopy does not necessarily provide additional benefit over standard WLC. However, the XBCM may provide better SE and a diagnostic advantage in instances of HG disease recurrence.

In vitro assessment of intra-operative and post-operative environment in reducing bladder cancer recurrence

Urinary bladder cancer is a common cancer worldwide. Currently, the modality of treating and monitoring bladder cancer is wide. Nonetheless, the high recurrence rate of non-muscle-invasive bladder cancer after surgical resection is still unsatisfactory. Hereby, our study demonstrated whether the intra-operative and post-operative environments will affect bladder cancer recurrence utilizing in vitro cell line model. Bladder cancer cell lines were submerged in four different irrigating fluids for assessing their tumorigenic properties. Our results showed that sterile water performed the best in terms of the magnitude of cytotoxicity to cell lines. Besides, we also investigated cytotoxic effects of the four irrigating agents as well as mitomycin C (MMC) in normothermic and hyperthermic conditions. We observed that sterile water and MMC had an increased cytotoxic effect to bladder cancer cell lines in hyperthermic conditions. Altogether, our results could be translated into clinical practice in the future by manipulating the intra-operative and post-operative conditions in order to lower the chance of residual cancer cells reimplant onto the bladder, which in turns, reducing the recurrence rate of bladder cancers.

Analysis of urinary kallikrein-related peptidase 13 for monitoring bladder cancer

Background Bladder cancer (BC) is one of the 10 most common types of cancer worldwide, with approximately 550,000 new cases each year. Early detection and appropriate diagnosis are important factors in successful treatment of the disease. Material and methods We used specific fluorogenic substrate for the quantitative determination of urine kallikrein 13 (KLK13) activity in healthy (n = 15) and BC (n = 54) patients. The proteolytic activity in individual urine samples was determined by fluorescence measurements. Then, immunoenzymatic analyses (ELISA, Western blot) were performed to confirm the presence of KLK13 in the tested samples. Results Urine samples from patients with G2 and G3 grade BC contained proteolytically active KLK13, as confirmed by kinetic analysis and immunochemical detection. KLK13 was not detected in the urine of patients with G1 grade BC. Discussion Previous clinical study reveals the KLK13 significance for BC prognosis as increased KLK13 expression was highlighted in bladder tumours compared to normal adjacent tissues. Our findings correlate to the report. KLK13 activity was confirmed in BC patients with G2 and G3 stage of disease development. Conclusions Using specific chromogenic/fluorogenic peptides could be useful for the non-invasive disease monitoring of BC progress.

Multiplexed detection of bladder cancer microRNAs based on core-shell-shell magnetic quantum dot microbeads and cascade signal amplification

Bladder Cancer (BC) is the tenth most common cancer worldwide and the most malignant disease of the urinary system. The development of effective biomarkers and detection methods is of great significance for BC diagnosis and monitoring. This study demonstrated a microscopic imaging method for multiplexed detecting BC microRNAs (miRNAs) based on core-shell-shell bifunctional magnetic fluorescent beads (MFBs), with the integration of catalytic hairpin assembly (CHA) and terminal deoxynucleotidyl transferase (TdT)-mediated DNA polymerization. Assembled with quantum dots and magnetic nanoparticles, MFBs were utilized as the dual-functional carriers for optical encoding and magnetic separation. By introducing CHA-TdT cascade amplification, three encoded MFBs presented distinct changes of emissive-color, which could be decoded by the common fluorescence microscope. The strategy could reach femtomolar level for the detection of single miRNA, showing a wide linear dynamic range (6 logs). Point-mutation discrimination capability was also achieved for precise miRNA analyses. Moreover, this MFB-CHA-TdT strategy was capable of detecting BC-related miRNAs in clinical serum specimens, showing a great consistence with qRT-PCR. Overall, this work enabled multiplexed detection of miRNAs with ideal sensitivity, specificity and accuracy, which could be applied to the clinical detection and early diagnosis of BC.