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1.
Methods Mol Biol ; 1660: 355-364, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28828671

RESUMO

Extracellular vesicles (EVs) are present in a variety of bodily fluids and they play an important role in cellular communications and signal transduction mechanisms. Studies have shown that the number of EVs and EV-associated biomarkers (i.e., proteins, nucleic acids and lipids) can be used to aid clinical diagnosis. Although ultracentrifugation is commonly used for EV isolation, it is not practical for clinical settings. Here, we developed an integrated double-filtration device that isolated and enriched EVs from urine, and subsequently detected/quantified EVs from urine via microchip ELISA. Results showed that the concentration of EVs was significantly elevated compared to healthy controls. Receiver operating characteristic analysis demonstrated that this integrated EV quantification device had a sensitivity of 81.3% at a specificity of 90% (16 bladder cancer patients and eight healthy controls). Thus, this integrated device shows great potential to supplement urine cytology for diagnosis of bladder cancer in point-of-care (POC) settings.


Assuntos
Vesículas Extracelulares/metabolismo , Filtração/instrumentação , Filtração/métodos , Dispositivos Lab-On-A-Chip , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/urina , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática/métodos , Vesículas Extracelulares/química , Humanos
2.
Sci Rep ; 7: 46224, 2017 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-28436447

RESUMO

Extracellular vesicles (EVs), including exosomes and microvesicles, are present in a variety of bodily fluids, and the concentration of these sub-cellular vesicles and their associated biomarkers (proteins, nucleic acids, and lipids) can be used to aid clinical diagnosis. Although ultracentrifugation is commonly used for isolation of EVs, it is highly time-consuming, labor-intensive and instrument-dependent for both research laboratories and clinical settings. Here, we developed an integrated double-filtration microfluidic device that isolated and enriched EVs with a size range of 30-200 nm from urine, and subsequently quantified the EVs via a microchip ELISA. Our results showed that the concentration of urinary EVs was significantly elevated in bladder cancer patients (n = 16) compared to healthy controls (n = 8). Receiver operating characteristic (ROC) analysis demonstrated that this integrated EV double-filtration device had a sensitivity of 81.3% at a specificity of 90% (16 bladder cancer patients and 8 healthy controls). Thus, this integrated device has great potential to be used in conjunction with urine cytology and cystoscopy to improve clinical diagnosis of bladder cancer in clinics and at point-of-care (POC) settings.


Assuntos
Vesículas Extracelulares/metabolismo , Filtração/instrumentação , Microfluídica/instrumentação , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/urina , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática , Vesículas Extracelulares/ultraestrutura , Humanos , Reprodutibilidade dos Testes , Reologia
3.
Expert Rev Mol Diagn ; 16(4): 449-59, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26777725

RESUMO

The striking prevalence of HIV, TB and malaria, as well as outbreaks of emerging infectious diseases, such as influenza A (H7N9), Ebola and MERS, poses great challenges for patient care in resource-limited settings (RLS). However, advanced diagnostic technologies cannot be implemented in RLS largely due to economic constraints. Simple and inexpensive point-of-care (POC) diagnostics, which rely less on environmental context and operator training, have thus been extensively studied to achieve early diagnosis and treatment monitoring in non-laboratory settings. Despite great input from material science, biomedical engineering and nanotechnology for developing POC diagnostics, significant technical challenges are yet to be overcome. Summarized here are the technical challenges associated with POC diagnostics from a RLS perspective and the latest advances in addressing these challenges are reviewed.


Assuntos
Doenças Transmissíveis/diagnóstico , Países em Desenvolvimento , Técnicas de Diagnóstico Molecular/métodos , Testes Imediatos/economia , Humanos , Técnicas de Diagnóstico Molecular/economia , Técnicas de Diagnóstico Molecular/normas , Testes Imediatos/normas
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