Point-of-care detection of extracellular vesicles: Sensitivity optimization and multiple-target detection

Oliveira-Rodríguez, Myriam; Serrano-Pertierra, Esther; Garcı́a, Agustı́n Costa; López-Martín, Sara; Yáñez-Mó, Marı́a; Cernuda-Morollón, Eva; Blanco‐López, M. Carmen

doi:10.1016/j.bios.2016.08.001

Cited by 90 publications

(63 citation statements)

References 35 publications

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“…The signal intensities were measured with the ESEQuant LR3 lateral flow strip reader (Qiagen, Madrid, Spain). For more details regarding the nanoparticle-antibody conjugation and the preparation of the immunostrip, see the studies previously published by Oliveira-Rodríguez et al [14,15]. …”

Section: Methodsmentioning

confidence: 99%

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Castro-Marrero

Serrano-Pertierra

Oliveira-Rodríguez

et al. 2018

J of Extracellular Vesicle

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Chronic Fatigue Syndrome (CFS), also known as Myalgic Encephalomyelitis (ME) is an acquired, complex and multisystem condition of unknown etiology, no established diagnostic lab tests and no universally FDA-approved drugs for treatment. CFS/ME is characterised by unexplicable disabling fatigue and is often also associated with numerous core symptoms. A growing body of evidence suggests that extracellular vesicles (EVs) play a role in cell-to-cell communication, and are involved in both physiological and pathological processes. To date, no data on EV biology in CFS/ME are as yet available. The aim of this study was to isolate and characterise blood-derived EVs in CFS/ME. Blood samples were collected from 10 Spanish CFS/ME patients and 5 matched healthy controls (HCs), and EVs were isolated from the serum using a polymer-based method. Their protein cargo, size distribution and concentration were measured by Western blot and nanoparticle tracking analysis. Furthermore, EVs were detected using a lateral flow immunoassay based on biomarkers CD9 and CD63. We found that the amount of EV-enriched fraction was significantly higher in CFS/ME subjects than in HCs (p = 0.007) and that EVs were significantly smaller in CFS/ME patients (p = 0.014). Circulating EVs could be an emerging tool for biomedical research in CFS/ME. These findings provide preliminary evidence that blood-derived EVs may distinguish CFS/ME patients from HCs. This will allow offer new opportunities and also may open a new door to identifying novel potential biomarkers and therapeutic approaches for the condition.

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Section: Methodsmentioning

confidence: 99%

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Castro-Marrero

Serrano-Pertierra

Oliveira-Rodríguez

et al. 2018

J of Extracellular Vesicle

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“…As the LFA implemented MAbs 1C4 and 2F8 against C. sakazakii serotypes O1 and O2 are characterized by a very high affinity (Scharinger et al, 2016 ), test improvement is not a matter of antibody optimization but rather of signal amplifying strategies (Urusov et al, 2016 ). For instance, it is possible to considerable increase intensity of test lines and thus reduce the detection range by using gold/silver nanoparticles (Raeisossadati et al, 2016 ; Oliveira-Rodriguez et al, 2017 ). An increased sensitivity might improve the applicability of the LFA for on-site analyses.…”

Section: Discussionmentioning

confidence: 99%

Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakii Serotypes O1 and O2

et al. 2017

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The ubiquitous and opportunistic pathogen Cronobacter sakazakii is responsible for severe meningitis, sepsis, and necrotizing enterocolitis in neonates and infants associated with ingestion of contaminated powdered infant formula (PIF). The current ISO method for isolation and detection of Cronobacter spp. is laborious, time-consuming and expensive. In this study, a multiplexed lateral flow test strip was developed to rapidly detect and simultaneously serotype O1 and O2 C. sakazakii serotypes. The assay is based on two monoclonal antibodies (MAb) that specifically bind to the lipopolysaccharides (LPS) of these pathogens. The test strip provides results very quickly; C. sakazakii could be detected in pure culture within 15 min with a sensitivity of 107 CFU/ml. After non-selective enrichment for 18 h as low as one Cronobacter cell per g PIF could be detected. Moreover, the established lateral flow assay (LFA) offers excellent specificity showing no cross-reactivity with other C. sakazakii serotypes, Cronobacter species or Enterobacteriaceae tested. These characteristics, together with several advantages such as speed, simplicity in performance, low analysis cost, and no requirement of specialized skills or sophisticated equipment make the developed multiplexed LFA suitable for reliable detection and serotyping of C. sakazakii serotypes O1 and O2.

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“…Many LFSAs utilize antibodies to detect specific biomolecules [ 29 , 31 , 32 , 33 ]. Before applying the sample to the strip, it must be mixed with detection antibodies that bind to the target molecule that is conjugated to a secondary molecule to provide an analytical output for qualitative analysis.…”

Section: Disease Detectionmentioning

confidence: 99%

“…The second line, or test line, contains immobilized antibodies that bind to the target molecule. Current LFSA results can be interpreted as a fluorescent signal present on the test line [ 32 ], as a visually present test line [ 29 , 30 , 33 ], or by measuring a pressure output from a chemical reaction [ 31 ].…”

Section: Disease Detectionmentioning

confidence: 99%

Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research

Campbell

Balhoff

Landwehr

et al. 2018

IJMS

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Recent developments in microfluidic devices, nanoparticle chemistry, fluorescent microscopy, and biochemical techniques such as genetic identification and antibody capture have provided easier and more sensitive platforms for detecting and diagnosing diseases as well as providing new fundamental insight into disease progression. These advancements have led to the development of new technology and assays capable of easy and early detection of pathogenicity as well as the enhancement of the drug discovery and development pipeline. While some studies have focused on treatment, many of these technologies have found initial success in laboratories as a precursor for clinical applications. This review highlights the current and future progress of microfluidic techniques geared toward the timely and inexpensive diagnosis of disease including technologies aimed at high-throughput single cell analysis for drug development. It also summarizes novel microfluidic approaches to characterize fundamental cellular behavior and heterogeneity.

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Point-of-care detection of extracellular vesicles: Sensitivity optimization and multiple-target detection

Cited by 90 publications

References 35 publications

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Circulating extracellular vesicles as potential biomarkers in chronic fatigue syndrome/myalgic encephalomyelitis: an exploratory pilot study

Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakii Serotypes O1 and O2

Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research

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