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J Breath Res ; 10(3): 036009, 2016 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-27447200


There are many respiratory infections such as influenza that cause epidemics. These respiratory infection epidemics can be effectively prevented by determining the presence or absence of infections in patients using frequent tests. We think that self-diagnosis may be possible using a system that can collect and detect biological aerosol particles in the patient's breath because breath sampling is easy work requiring no examiner. In this paper, we report a sensing system for biological aerosol particles (SSBAP) with a disposable device. Using the system and the device, someone with no medical knowledge or skills can safely, easily, and rapidly detect infectious biological aerosol particles. The disposable device, which is the core of the SSBAP, can be an impactor for biological aerosol particles, a flow-cell for reagents, and an optical window for the fluorescent detection of collected particles. Furthermore, to detect the fluorescence of very small collected particles, this disposable device is covered with a light-blocking film that lets only fluorescence of particles pass through a fluorescence detector of the SSBAP. The SSBAP using the device can automatically detect biological aerosol particles by the following process: collecting biological aerosol particles from a patient's breath in a sampling bag by the impaction method, labeling the collected biological aerosol particles with fluorescent dyes by the antigen-antibody reaction, removing free fluorescent dyes, and detecting the fluorescence of the biological aerosol particles. The collection efficiency of the device for microspheres aerosolized in the sampling bag was more than 97%, and the SSBAP with the device could detect more than 8.3 × 10(3) particles l(-1) of aerosolized influenza virus particles within 10 min.

Aerossóis/análise , Poluição do Ar/análise , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Microbiologia Ambiental , Testes Respiratórios , Fluorescência , Humanos , Manejo de Espécimes
J Phys Chem B ; 115(4): 653-9, 2011 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-21162543


The calcium hydroxyapatite Ca(10)(PO(4))(6)(OH)(2) (Hap) nanoparticles were prepared by using microreactor and employed these Hap nanoparticles to clarify the adsorption behavior of proteins. The size of Hap particles produced by the microreactor reduced in the order of a hardness of the reaction conditions for mixing Ca(OH)(2) and H(3)PO(4) aqueous solutions, such as flow rates of both solutions and temperature. Finally, the size of the smallest Hap nanoparticle became 2 × 15 nm(2), similar to that of BSA molecule (4 × 14 nm(2)). It is noteworthy that the smallest Hap nanoparticles still possesses rodlike shape, suggesting that particles are grown along c-axis even though the reactants mixed very rapidly in narrow channels of the microreactors. The X-ray diffraction patterns of the Hap nanoparticles revealed that the crystallinity of the materials produced by the microreactor is low. The FTIR measurement indicated that the Hap nanoparticles produced by microreactor were carbonate-substituted type B Hap, where the carbonate ions replace the phosphate ions in the crystal lattice. All the adsorption isotherms of acidic bovine serum albumin (BSA), neutral myoglobin (MGB), and basic lysozyme (LSZ) onto Hap nanoparticles from 1 × 10(-4) mol/dm(3) KCl solution were the Langmuirian type. The saturated amounts of adsorbed BSA (n(S)(BSA)) for the Hap nanoparticles produced by microreactor were decreased with decrease in the mean particle length, and finally it reduced to zero for the smallest Hap nanoparticles. Similar results were observed for the adsorption of LSZ; the saturated amounts of adsorbed LSZ (n(S)(LSZ)) also reduced to zero for the smallest Hap nanoparticles. However, in the case of MGB, the saturated mounts of adsorbed MGB (n(S)(MGB)) are also depressed with decreased in their particle size, but about half of MGB molecules still adsorbed onto the smallest Hap nanoparticles. This difference in the protein adsorption behavior was explained by the difference in the size and flexibility of three kinds of proteins. The reduction of n(S)(BSA) is due to the decrease in the fraction of C sites on the side face of each Hap nanoparticle; i.e., there is not enough area left on the nanoparticle surface to adsorb large BSA molecules even though the BSA molecules are soft and their conformations are alterable. The reduction of n(S)(LSZ) was explained by the reduction of P sites. Further, rigidity of the LSZ molecules was given another possibility of the depression of n(S)(LSZ) for the Hap nanoparticles. However, MGB molecules with small and soft structure were adsorbed on the Hap nanoparticle surface by changing their conformation. We could control the amounts of adsorbed proteins by changing the particle size of Hap in the nanometer range and kinds of proteins. These obtained results may be useful for developing biomimetic materials for bone grafts and successful surgical devices in the biochemical field.

Tecnologia Biomédica/métodos , Durapatita/síntese química , Nanopartículas/química , Proteínas/isolamento & purificação , Adsorção , Tecnologia Biomédica/instrumentação , Durapatita/química , Desenho de Equipamento , Microscopia Eletrônica de Transmissão , Muramidase/isolamento & purificação , Mioglobina/isolamento & purificação , Conformação Proteica , Soroalbumina Bovina/isolamento & purificação , Propriedades de Superfície
Biosens Bioelectron ; 24(11): 3299-305, 2009 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-19450964


A semi-automated bacterial spore detection system (BSDS) was developed to detect biological threat agents (e.g., Bacillus anthracis) on-site. The system comprised an aerosol sampler, micro-fluidic chip-A (for spore germination and cell lysis), micro-fluidic chip-B (for extraction and detection of genomic DNA) and an analyzer. An aerosol with bacterial spores was first collected in the collection chamber of chip-A with a velocity of 300 l/min, and the chip-A was taken off from the aerosol sampler and loaded into the analyzer. Reagents packaged in the chip-A were sequentially applied into the chamber. The genomic DNA extract from spore lyzate was manually transferred from chip-A to chip-B and loaded into the analyzer. Genomic DNA in chip-B was first trapped on a glass bead column, washed with various reagents, and eluted to the detection chamber by sequential auto-dispensing. Isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) with fluorescent measurement was adopted to amplify and detect target DNA. Bacillus subtilis was the stimulant of biological warfare agent in this experiment. Pretreatment conditions were optimized by examining bacterial target DNA recovery in the respective steps (aerosol collection, spore germination, cell lysis, and DNA extraction), by an off-chip experiment using a real-time polymerase chain reaction quantification method. Without the germination step, B. subtilis spores did not demonstrate amplification of target DNA. The detection of 10(4) spores was achieved within 2h throughout the micro-fluidic process.

Aerossóis/análise , Bacillus anthracis/genética , Bacillus anthracis/isolamento & purificação , Técnicas Biossensoriais/instrumentação , DNA Bacteriano/análise , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Microbiologia do Ar , Algoritmos , DNA Bacteriano/genética , Monitoramento Ambiental/instrumentação , Monitoramento Ambiental/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Técnicas de Amplificação de Ácido Nucleico/métodos , Esporos Bacterianos/genética , Esporos Bacterianos/isolamento & purificação