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Exosomes are cell-derived membrane-enclosed biological nanoparticles that carry lots of parental molecular information, and are recognized as an ideal biomarker for non-invasive diagnosis. However, due to the low abundance of exosomes in plasma samples and the interferences from complex biological matrices, the sensitive and direct detection of exosomes still remains a challenge. Here, by combining the direct magnetic isolation with in situ fluorescence imaging, we developed a Single Microbead-based Fluorescent Aptasensor (SMFA) for specific enrichment and sensitive quantification of exosomes from plasma. In the SMFA, a single aptamer-modified microbead (MB) served as the reaction carrier so that the specific exosomes inserted with a fluorescent anchor will be highly enriched on the single MB. By in situ fluorescence imaging to monitor the fluorescence signals on the single MB, sensitive detection of exosomes can be realized without the requirement of any signal amplification routes, and as low as 4.9 × 104 particles per μL of exosomes could be simply detected. More importantly, the SMFA could be applied for direct detection of the exosomes from small amounts of clinical plasma samples without prior purification procedures, indicating its great potential applications in clinical diagnostics.

Tumor-derived exosomes carrying unique surface proteins have shown great promise as novel biomarkers for liquid biopsies. However, point-of-care analysis for tumor-derived exosomes in the blood with low-cost and easy processing is still challenging. Herein, we develop an integrated approach, homogenous magneto-fluorescent exosome (hMFEX) nanosensor, for rapid and on-site tumor-derived exosomes analysis. Tumor-derived exosomes are captured immunomagnetically, which further initiates the aptamer-triggered assembly of DNA three-way junctions in homogenous solution containing aggregation-induced emission luminogens and graphene oxide, resulting in an amplified fluorescence signal. By integrating magnetic isolation and enhanced fluorescence measurement, the hMFEX nanosensor detects tumor-derived exosomes in the dynamic range spanning 5 orders of magnitude with high specificity, and the limit of detection is 6.56 × 104 particles/μL. Analyzing tumor-derived exosomes in limited volume plasma from breast cancer patients demonstrates the excellent clinical diagnostic efficacy of the hMFEX nanosensor. This study provides new insights into the point-of-care testing of tumor-derived exosomes for cancer diagnostics.

The detection of a low concentration of biomarkers required for an early diagnosis remains a tricky task to pursue. In this context, magnetic nanoparticles (MNPs) have been widely used to isolate/concentrate diluted samples. However, by applying the magnetic field, all MNPs are isolated, both those conjugated with the analyte and free MNPs. This excess of MNP can interfere with the analyte quantification. Herein, we propose a novel and versatile approach that ensures the selective isolation of the analyte-conjugated MNP by exploiting the electrophoretic and magnetophoretic behavior of respectively charged silica nanoparticles (SiNPs) and MNPs. In this work, the robustness and sensitivity of the developed isolation and fluorescence-based detection procedure were confirmed by comparing them with the standard method based on simple magnetic isolation of the analyte. The limit of detection (LOD) is improved by two orders of magnitude, namely, from 1 nM to 10 pM. This finding confirms the potentiality of the method and its possible application in the development of rapid and easy-to-use point-of-care devices.

Methicillin-resistant Staphylococcus aureus (MRSA) is considered to be one of the important hospital-acquired pathogens. MRSA is also commonly associated with hospital-acquired infections and mortality. Quantitative and precise detection of MRSA is essential for rapid diagnosis and subsequent effective disease management strategies. We herein developed a highly specific method for rapid MRSA detection that combines surface-enhanced Raman spectroscopy (SERS) nanotags and polymerase chain reaction (PCR). SERS provided the sensitivity and spectral multiplexing capability while PCR provided the specificity required for the assay. The method was tested by the simultaneous detection of two MRSA specific genes (mecA and femA) amplified from genomic DNA isolated from clinical specimens. Magnetic isolation and rapid duplex detection were performed to obtain a detectable signal down to 104 input copies within 80 min. This demonstrated the potential of the SERS-PCR based approach for the accurate identification of MRSA at an early-diagnosis stage. This study also provides an alternative approach to the existing methods for detecting clinical targets without compromising sensitivity and selectivity, and with minimal handling steps. We thus believe that this approach will find a broad application in clinical applications.

Grain boundary diffusion process (GBDP) with low melting-point alloy ribbons, namely Pr70Cu15Al15 (PCA), Pr65Tb5Cu15Al15 (PTCA-5) and Pr50Tb20Cu15Al15 (PTCA-20, at%), was applied to Nd-Fe-B sintered magnet (52 M) as thick as 6 mm. The coercivities of diffused magnets by PCA, PTCA-5 and PTCA-20 increase significantly to 19.6 kOe, 24.1 kOe and 24.3 kOe, respectively, from original 14.7 kOe. Microstructural investigations indicate that the coercivity enhancement of PCA magnet is mainly attributed to magnetic isolation by continuous Nd-rich grain boundary phases (GBP). However, the coercivity enhancement of PTCA-20 magnet is mainly owing to magnetic hardening by Tb-rich shells (TRS), and that of PTCA-5 magnet results from a combined strengthening of magnetic isolation by GBP and magnetic hardening by TRS. Meanwhile, the coercivity of PTCA-5 magnet is as high as that of PTCA-20 magnet, but the former consumes Tb only 0.60 wt%, lower than 1.23 wt% for the latter.

Background Highly efficient capture and detection of circulating tumor cells (CTCs) remain elusive mainly because of their extremely low concentration in patients’ peripheral blood. Methods We present an approach for the simultaneous capturing, isolation, and detection of CTCs using an immuno-fluorescent magnetic nanobead system (iFMNS) coated with a monoclonal anti-EpCAM antibody. Results The developed antibody nanobead system allows magnetic isolation and fluorescent-based quantification of CTCs. The expression of EpCAM on the surface of captured CTCs could be directly visualized without additional immune-fluorescent labeling. Our approach is shown to result in a 70–95% capture efficiency of CTCs, and 95% of the captured cells remain viable. Using our approach, the isolated cells could be directly used for culture, reverse transcription-polymerase chain reaction (RT-PCR), and immunocytochemistry (ICC) identification. We applied iFMNS for testing CTCs in peripheral blood samples from a lung cancer patient. Conclusions It is suggested that our iFMNS approach would be a promising tool for CTCs enrichment and detection in one step. Graphic abstract

Isolating and analyzing tumor-derived exosomes (TEX) can provide important information about the state of a tumor, facilitating early diagnosis and prognosis. Since current isolation methods are mostly laborious and expensive, we propose herein a fast and cost-effective method based on a magnetic nanoplatform to isolate TEX. In this work, we have tested our method using three magnetic nanostructures: (i) Ni magnetic nanowires (MNWs) (1500 &times, 40 nm), (ii) Fe3O4 nanorods (NRs) (41 &times, 7 nm), and (iii) Fe3O4 cube-octahedral magnetosomes (MGs) (45 nm) obtained from magnetotactic bacteria. The magnetic response of these nanostructures has been characterized, and we have followed their internalization inside canine osteosarcoma OSCA-8 cells. An overall depiction has been obtained using a combination of Fluorescence and Scanning Electron Microscopies. In addition, Transmission Electron Microscopy images have shown that the nanostructures, with different signs of degradation, ended up being incorporated in endosomal compartments inside the cells. Small intra-endosomal vesicles that could be precursors for TEX have also been identified. Finally, TEX have been isolated using our magnetic isolation method and analyzed with a Nanoparticle tracking analyzer (NanoSight). We observed that the amount and purity of TEX isolated magnetically with MNWs was higher than with NRs and MGs, and they were close to the results obtained using conventional non-magnetic isolation methods.

Human papillomavirus (HPV) and human immunodeficiency virus (HIV) are closely related infections; both diseases are mutually reinforcing due to the co-infection effects, which increase their clinical and epidemiological implications, raising urgent public health concerns. In the present study, a nano-system was developed for simultaneous detection of HPV and HIV targeting E6-HPV16 and Gag-HIV genes respectively. The target sequences from the samples of the patients suffering from head and neck cancer were isolated by magnetic glass particles (MGPs) and subsequently were hybridized with a mixture of the complementary probes attached to cadmium telluride quantum dots (CdTe QDs). Finally, the specific and successful binding was determined by an easy fluorescence analyses and was corroborated by PCR and electrochemistry. Red and green CdTe QDs were used for the dual-color detection of HPV and HIV respectively. Our nano-system can be applied as a proficient tool for the identification of HPV, HIV or both diseases simultaneously, getting a successful and high sensitivity system of detection.

Metallothioneins (MTs) are involved in heavy metal detoxification in a wide range of living organisms. Currently, it is well known that MTs play substantial role in many pathophysiological processes, including carcinogenesis, and they can serve as diagnostic biomarkers. In order to increase the applicability of MT in cancer diagnostics, an easy-to-use and rapid method for its detection is required. Hence, the aim of this study was to develop a fully automated and high-throughput assay for the estimation of MT levels. Here, we report the optimal conditions for the isolation of MTs from rabbit liver and their characterization using MALDI-TOF MS. In addition, we described a two-step assay, which started with an isolation of the protein using functionalized paramagnetic particles and finished with their electrochemical analysis. The designed easy-to-use, cost-effective, error-free and fully automated procedure for the isolation of MT coupled with a simple analytical detection method can provide a prototype for the construction of a diagnostic instrument, which would be appropriate for the monitoring of carcinogenesis or MT-related chemoresistance of tumors.

Iron-based hierarchical nanostructures with enhanced properties have attracted intense interest as sorbents for water treatment. It is important to design a facile and scalable procedure for the preparation of absorbents with hierarchical structures. Here, a green solid-state method for the fabrication of Fe/FeOOH hierarchical nanostructures (Fe-HNs) is developed. The obtained samples with sizes in the range of 500–800 nm are three-dimensional (3D) hierarchical structures assembled from ultrathin nanosheets. A plausible “reduction–oxidation” mechanism is proposed for the formation process of the Fe-HNs. The as-prepared Fe-HNs are utilized as an adsorbent for the removal of Congo red and Cr(VI) ions from aqueous solution. The results show that the adsorbent exhibits high adsorption capacity, fast adsorption rate, and superior magnetic isolation. These Fe-HNs prepared by a facile solid-state method display very promising potential application in water treatment as an adsorbent.

The authors describe a method for the colorimetric determination of unamplified microRNA. It is based on the use of citrate-capped gold nanoparticles (AuNPs) and, alternatively, a microRNA-probe hybrid or a magnetically extracted microRNA that serve as stabilizers against the salt-induced aggregation of AuNPs. The absorbance ratios A525/A625 of the reacted AuNP solutions were used to quantify the amount of microRNA. The assay works in the range of 5–25 pmol microRNA. The lower limit of detection (LOD) is 10 pmol. The performance of the method was tested by detection of microRNA-210-3p in totally extracted urinary microRNA from normal, benign, and bladder cancer subjects. The sensitivity and specificity for qualitative detection of urinary microRNA-210-3p using the assay are 74% and 88% respectively, which is consistent with real time PCR based assays. The assay was applied to the determination of specific microRNA by using its specific oligo targeter or following magnetic isolation of the desired microRNA. The method is simple, cost-efficient, has a short turn-around time and requires minimal equipment and personnel.

Extracellular vesicles (EVs) can mediate intercellular communication by transferring cargo proteins and nucleic acids between cells. The pathophysiological roles and clinical value of EVs are under intense investigation, yet most studies are limited by technical challenges in the isolation of nanoscale EVs (nEVs). Here, we report a lipid nanoprobe that enables spontaneous labelling and magnetic enrichment of nEVs in 15 minutes, with isolation efficiency and cargo composition similar to what can be achieved by the much slower and bulkier method of ultracentrifugation. We also show that the lipid nanoprobes, which allow for downstream analyses of nucleic acids and proteins, enabled the identification of EGFR and KRAS mutations following nEV isolation from blood plasma from non-small-cell lung-cancer patients. The efficiency and versatility of the lipid nanoprobe opens up opportunities in point-of-care cancer diagnostics.

The long-term aim of this work is to develop a biosensing system that rapidly detects bacterial targets of interest, such as Escherichia coli, in drinking and recreational water quality monitoring. For these applications, a standard sample size is 100 mL, which is quite large for magnetic separation microfluidic analysis platforms that typically function with &lt, 20 &micro, L/s throughput. Here, we report the use of 1.5-&micro, m-diameter magnetic microdisc to selectively tag target bacteria, and a high-throughput microfluidic device that can potentially isolate the magnetically tagged bacteria from 100 mL water samples in less than 15 min. Simulations and experiments show ~90% capture efficiencies of magnetic particles at flow rates up to 120 &micro, L/s. Also, the platform enables the magnetic microdiscs/bacteria conjugates to be directly imaged, providing a path for quantitative assay.

A significant coercivity enhancement of the commercial NdFeB magnets with the magnetic properties of (BH)max = 48.4 MGOe and iHc = 17.5 kOe through grain boundary diffusion (GBD) with low-melting Tb55R20Cu25 alloys is demonstrated. Adopting Tb55R20Cu25 alloys as GBD sources is effective in increasing coercivity to 29.0 kOe for R = None, 23.8 kOe for R = Y, 25.6 kOe for R = La, 28.0 kOe for R = Ce, respectively. Yet, (BH)max is slightly reduced to 46.2-48.2 MGOe. The preferential appearance of Cu at grain boundary and triple junction of the grains, and the core-shell structure occurred due to Tb at grain surface remarkably enhance the coercivity. Interestingly, higher coercivity enhancement per wt% Tb usage (ΔiHc/wt%Tb) of 7.2 kOe/wt% for the magnet with Tb55Ce25Cu25 than 5.9 kOe/wt% for that with Tb75Cu25 has been found due to the magnetic isolation effect caused by the preferential appearance of Ce at grain boundary, though a slight lower coercivity enhancement was found for the samples with R = Y and La. Lower melting point (637 °C) for Tb55Ce20Cu25 than Tb75Cu25 (743 °C) leads to larger diffusion depth of Tb into the magnet and therefore contributes to higher efficiency of coercivity enhancement for the magnet with R=Ce.

Ce-based R2Fe14B (R= rare-earth) nano-structured permanent magnets consisting of (Ce,Nd)2Fe14B core-shell grains separated by a non-magnetic grain boundary phase, in which the relative amount of Nd to Ce is higher in the shell of the magnetic grain than in its core, were fabricated by Nd-Cu infiltration into (Ce,Nd)2Fe14B hot-deformed magnets. The coercivity values of infiltrated core-shell structured magnets are superior to those of as-hot-deformed magnets with the same overall Nd content. This is attributed to the higher value of magnetocrystalline anisotropy of the shell phase in the core-shell structured infiltrated magnets compared to the homogeneous R2Fe14B grains of the as-hot-deformed magnets, and to magnetic isolation of R2Fe14B grains by the infiltrated grain boundary phase. First order reversal curve (FORC) diagrams suggest that the higher anisotropy shell suppresses initial magnetization reversal at the edges and corners of the R2Fe14B grains.

The majority of carcinomas that were developed due to the infection with human papillomavirus (HPV) are caused by high-risk HPV types, HPV16 and HPV18. These HPV types contain the E6 and E7 oncogenes, so the fast detection of these oncogenes is an important point to avoid the development of cancer. Many different HPV tests are available to detect the presence of HPV in biological samples. The aim of this study was to design a fast and low cost method for HPV identification employing magnetic isolation, polymerase chain reaction (PCR) and electrochemical detection. These assays were developed to detect the interactions between E6-HPV16 oncogene and magnetizable particles (MPs) using commercial Dynabeads M-280 Streptavidin particles and laboratory-synthesized “homemade” particles called MANs (MAN-37, MAN-127 and MAN-164). The yields of PCR amplification of E6-HPV16 oncogene bound on the particles and after the elution from the particles were compared. A highest yield of E6-HPV16 DNA isolation was obtained with both MPs particles commercial M-280 Streptavidin and MAN-37 due to reducing of the interferents compared with the standard PCR method. A biosensor employing the isolation of E6-HPV16 oncogene with MPs particles followed by its electrochemical detection can be a very effective technique for HPV identification, providing simple, sensitive and cost-effective analysis. Většina karcinomů, které byly vyvolané v důsledku infekce lidským papilomavirem (HPV), jsou způsobeny vysoce rizikovými typy HPV, HPV16 a HPV18. Tyto typy HPV obsahují E6 a E7 onkogeny, takže rychlá detekce těchto onkogenů je důležitý bod k zabránění rozvoje rakoviny. Mnoho různých HPV testů je k dispozici pro detekci přítomnosti HPV v biologických vzorcích. Cílem této studie bylo navrhnout rychlou a levnou metodu pro identifikaci HPV využívající magnetickou izolaci, polymerázovou řetězovou reakci (PCR) a elektrochemickou detekci. Tyto testy byly vyvinuty s cílem odhalit interakce mezi E6-HPV16 onkogenem a magnetizovatelnými částicemi (MPS) s použitím komerčních částic Dynabeads M-280 Streptavidin a laboratorně syntetizovaných "homemade" částic zvaných MAN (MAN-37, MAN-127 a MAN-164). Byly porovnány výnosy z PCR amplifikace onkogenu E6-HPV16 vázaného na částicích a po eluci z částic. Nejvyšším výnos z izolace E6-HPV16 DNA byl získán s oběma komerčními MPs částicemi M-280 Streptavidin a MAN-37 v důsledku snížení interferencí ve srovnání se standardní metodou PCR. Biosenzor používající izolaci E6-HPV16 onkogenu s MPs částicemi následovaný jeho elektrochemickou detekci může být velmi účinná technika pro identifikaci HPV, která umožňuje jednoduchou, citlivou a nákladově efektivní analýzu.