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1.
J Mech Behav Biomed Mater ; 150: 106156, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38041883

RESUMO

INTRODUCTION: To develop a calcium silicate (CaSi)-based cement containing dimethyl sulfoxide (DMSO) and cement deliver device for new root canal filling technique, and to assess the flow behavior, leakage, and root canal filling quality of CaSi containing DMSO. METHODS: CaSi containing DMSO (CSC-DMSO) and CaSi containing PEG (CSC-PEG) were prepared, and the flow characteristics of both cements were compared in gypsum and resin channels using a high-speed camera. Eight root canals were obturated by CSC-DMSO or CSC-PEG using a cement delivery device, and root canal filling quality was assessed in terms of filling length using periapical radiographs. The filling length was evaluated by 'apico-coronal extension,' measuring length in reference to apical constriction. Microleakage was measured for thirty human molars that were randomly filled with CSC-DMSO, CSC-PEG, or gutta-percha and AH plus. Preliminary obturation of CSC-DMSO with cement delivery device in human teeth was analyzed in terms of filling length and void, using periapical radiographs. Statistical analysis was performed with the Kruskal Wallis test for simulated root canal fillings and one-way ANOVA for leakage test. RESULTS: The flow speed of CSC-DMSO reduced in gypsum channels compared to resin channels, but CSC-PEG did not exhibit significant differences in the channels. The median absolute value of apico-coronal extension was significantly lower in CSC-DMSO compared to CSC-PEG (p < 0.05). Microleakage did not statistically differ between the groups (p > 0.05). In the preliminary obturation, the mean apico-coronal extension of CSC-DMSO was -0.297 ± 0.724 mm, while CSC-PEG was not feasible due to excess apical extrusions. CONCLUSIONS: CSC-DMSO could be considered as an alternative filling material for root canal obturation.


Assuntos
Materiais Restauradores do Canal Radicular , Humanos , Materiais Restauradores do Canal Radicular/farmacologia , Dimetil Sulfóxido , Cavidade Pulpar , Cálcio , Sulfato de Cálcio , Cimentos de Ionômeros de Vidro , Resinas Epóxi
2.
Int Endod J ; 56(6): 765-774, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36825362

RESUMO

AIM: To evaluate the efficacy of a novel ultrasonic irrigation device, remotely-generated irrigation with a non-invasive sound field enhancement (RINSE) system, in removing biofilm-mimicking hydrogel from a simulated isthmus model and compare it with sonically- and ultrasonically-activated irrigation systems. METHODOLOGY: A polycarbonate root canal model containing two standardized root canals (apical diameter of 0.20 mm, 4% taper, 18 mm long with a coronal reservoir) connected by three isthmuses (0.40 mm deep, 2 mm high, 4 mm long) was used as the test model. The isthmuses were filled with a hydroxyapatite powder-containing hydrogel. The canals were filled with irrigant, and the models were randomly assigned to the following activation groups (n = 15): EndoActivator (EA), ultrasonically activated irrigation (UAI), and RINSE system (RS). Syringe irrigation (SI) with a 30G needle served as the control. Standardized images of the isthmuses were taken before and after irrigation, and the amount of hydrogel removed was determined using image analysis software and compared across groups using anova (p < .05). RESULTS: Hydrogel removal was significantly higher with the RS (83.7%) than with UAI, EA, or SI (p ≤ .01). UAI (69.2%) removed significantly more hydrogel than SI and EA (p < .05), while there was no significant difference between SI (24.3%) and EA (25.7%) (p = .978). CONCLUSIONS: RINSE system resulted in the most hydrogel removal, performing better than UAI or EA. The effect of RS was also not reliant on the insert or tip entering the pulp chamber or root canal, making it particularly useful in conservative endodontics.


Assuntos
Cavidade Pulpar , Tratamento do Canal Radicular , Ondas Ultrassônicas , Biofilmes/efeitos da radiação , Cavidade Pulpar/microbiologia , Cavidade Pulpar/efeitos da radiação , Hidrogéis , Irrigantes do Canal Radicular , Preparo de Canal Radicular , Hipoclorito de Sódio , Irrigação Terapêutica/métodos , Modelos Anatômicos , Tratamento do Canal Radicular/instrumentação , Tratamento do Canal Radicular/métodos
3.
Ultrason Sonochem ; 90: 106168, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36137469

RESUMO

Root canal treatment is performed to remove the bacteria proliferating in the root canals of a tooth. Many conventional root canal irrigation methods use an instrument inserted into the root canals. However, bacteria removal is often incomplete in the apical region of the root canal, and the treatment carries clinical risks, such as instrument fracture and extrusion of irrigation liquid through the canal apex. We here suggest a novel, remotely generated high-intensity ultrasound irrigation system that exhibits better irrigation performance and a reduced clinical risk. Our device employs powerful ultrasonic waves generated by a transducer placed outside a target tooth. The generated ultrasonic waves are guided to travel into the root canals. In the root canals of the target tooth, acoustic cavitation occurs, and vapor bubbles are created. The dynamic motions of vapor bubbles create remarkable cleaning effects. Using root canal models, we tested the cleaning performance of the proposed system and compared it with other conventional irrigation methods. The results revealed that biofilm in the apical region of the root canal models can be removed exclusively using the proposed system, thus demonstrating an improvement in cleaning performance. We also measured pressure at the apex of the root canals of an extracted tooth while operating the proposed system. Our system exhibited a smaller pressure compared to the syringe irrigation method, thus suggesting a reduced risk of apical extrusion of the irrigation liquid. Since the proposed system operates without inserting instruments into the root canal, it can clean multiple root canals in a tooth simultaneously with a single treatment. The proposed device would be a breakthrough in root canal treatment in terms of irrigation performance, clinical safety, and ease of treatment.


Assuntos
Irrigantes do Canal Radicular , Preparo de Canal Radicular , Preparo de Canal Radicular/métodos , Ápice Dentário , Cavidade Pulpar , Irrigação Terapêutica/métodos , Hipoclorito de Sódio
4.
ACS Appl Mater Interfaces ; 13(38): 46015-46021, 2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34544236

RESUMO

We propose a novel osmotic soft valve consisting of an osmosis membrane and hydrogel films. In our osmotic valve system, material selectivity is determined by the osmosis membrane, and the hydrogel film, which deforms depending on the ion concentration of the surrounding solution, controls the passage area of the membrane. Independently controlling the material selectivity and permeability allowed us to design an osmotic soft valve with an osmotic flow rate that increases with osmotic pressure at low pressures but decreases with osmotic pressure at high pressures. We demonstrate a representative application of our hydrogel valve system in a portable power generator utilizing reverse electrodialysis (RED). As the permeability varied with concentration, the hydrogel valve was able to maintain the electric power of the RED for 30 min with only an ∼10% change. Our study provides techniques to build osmotic soft valves that can serve as gating membranes in various osmosis and dialysis systems.

5.
Ultrason Sonochem ; 76: 105618, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34119904

RESUMO

The dynamic actions of cavitation bubbles in ultrasonic fields can clean surfaces. Gas and vapor cavitation bubbles exhibit different dynamic behaviors in ultrasonic fields, yet little attention has been given to the distinctive cleaning effects of gas and vapor bubbles. We present an experimental investigation of surface cleaning by gas and vapor bubbles in an ultrasonic field. Using high-speed videography, we found that the primary motions of gas and vapor bubbles responsible for surface cleaning differ. Our cleaning tests under different contamination conditions in terms of contaminant adhesion strength and surface wettability reveal that vapor and gas bubbles are more effective at removing contaminants with strong and weak adhesion, respectively, and furthermore that hydrophobic substrates are better cleaned by vapor bubbles. Our study not only provides a better physical understanding of the ultrasonic cleaning process, but also proposes novel techniques to improve ultrasonic cleaning by selectively employing gas and vapor bubbles depending on the characteristics of the surface to be cleaned.

6.
Proc Natl Acad Sci U S A ; 118(3)2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33431685

RESUMO

Mud nests built by swallows (Hirundinidae) and phoebes (Sayornis) are stable granular piles attached to cliffs, walls, or ceilings. Although these birds have been observed to mix saliva with incohesive mud granules, how such biopolymer solutions provide the nest with sufficient strength to support the weight of the residents as well as its own remains elusive. Here, we elucidate the mechanism of strong granular cohesion by the viscoelastic paste of bird saliva through a combination of theoretical analysis and experimental measurements in both natural and artificial nests. Our mathematical model considering the mechanics of mud nest construction allows us to explain the biological observation that all mud-nesting bird species should be lightweight.


Assuntos
Comportamento de Nidação/fisiologia , Passeriformes/fisiologia , Saliva/fisiologia , Andorinhas/fisiologia , Animais , Modelos Teóricos
7.
Sci Rep ; 10(1): 14731, 2020 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-32895450

RESUMO

In the chemical mechanical polishing process of semiconductor manufacturing, the concentration of 'large' particles ([Formula: see text]0.5 µm) in the slurry, which is considerably larger in size than the main abrasives ([Formula: see text] 0.1 µm), is a critical parameter that strongly influences manufacturing defects, yields, and reliabilities of large-scale-integrated circuits. Various instruments, so-called particle counters, based on light scattering, light extinction, and holography techniques have been developed to measure and monitor the large particle concentration in semiconductor fabs in real time. However, sizeable fluctuation in the measured particle concentration complicates the statistical process control in the fabs worldwide. Here, we show that an inherent fluctuation exists in the counting of large particles, which is universal, independent of instrument type, and quantitatively determined by the instrument's operation parameters. We analytically derive a statistical theory of the fluctuation based on Poisson statistics and validate the theory through experiments and Monte-Carlo simulation. Furthermore, we provide a strategy to enhance the measurement accuracy by statistically adjusting the instrumental parameters commonly involved in the particle counters. The present results and analyses could be useful for statistical process control in semiconductor fabs to prevent large particle-induced defects such as micro-scratches and pits on wafers.

8.
PLoS One ; 15(8): e0237848, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32817715

RESUMO

Dust masks are widely used to prevent the inhalation of particulate matter into the human respiratory organs in polluted air environments. The filter of a dust mask inherently obstructs the natural respiratory air flows, and this flow resistance is mainly responsible for the discomfort experienced when wearing a dust mask. In atmospheric conditions seriously contaminated with fine dust, it is recommended that common citizens wear a dust mask in their everyday lives, yet many people are reluctant to wear a dust mask owing to the discomfort experienced when wearing it for a long time. Understanding of physical reasons for the discomfort is thus crucial in designing a dust mask, but remains far from clear. This study presents a technique to quantify the wearing comfort of dust masks. By developing a respiration simulator to measure the pressure loss across a dust mask, we assessed the energy costs to overcome flow resistance when breathing through various types of dust masks. The energy cost for a single inhalation varies with the mask type in a range between 0 and 10 mJ. We compared the results with the survey results of 40 people about the wearing comfort of the dust masks, which revealed that the wearing comfort crucially depends on the energy cost required for air inhalation though the dust mask. Using the measured energy cost during inhalation as a parameter to quantify the wearing comfort, we present a comprehensive evaluation of the performance of dust masks in terms of not only the filtering performance but also the wearing comfort. Our study suggests some design principles for dust mask filters, auxiliary electric fans, and check valves.


Assuntos
Inalação/fisiologia , Máscaras/normas , Respiração , Dispositivos de Proteção Respiratória/normas , Adulto , Poeira/prevenção & controle , Orelha/fisiologia , Feminino , Filtração/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Material Particulado
9.
Soft Matter ; 16(24): 5571-5576, 2020 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-32542282

RESUMO

We construct a theoretical framework to understand the crack density of bloodstains by modeling whole blood as a suspension of binary size colloid particles. Our analysis based upon theories of soft capillarity and porous flows explains the observed increase of the crack density with increase of blood viscosity and decrease of environmental humidity. The results have direct implications on forensic science and medical diagnosis.


Assuntos
Manchas de Sangue , Modelos Teóricos , Viscosidade Sanguínea , Coloides , Humanos , Umidade
10.
Phys Rev E ; 101(4-1): 043108, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32422796

RESUMO

When a water drop impinges on a flat superhydrophobic surface, it bounces off the surface after a certain dwelling time, which is determined by the Rayleigh inertial-capillary timescale. Recent works have demonstrated that this dwelling time (i.e., contact time) is modified on curved superhydrophobic surfaces, as the drop asymmetrically spreads over the surface. However, the contact time on the curved surfaces still remains poorly understood, while no successful physical model for the contact time has been proposed. Here, we propose that the asymmetric spreading on the curved surface is driven by either the Coanda effect or inertia depending on the ratio of the drop diameter to the curvature diameter. Then, based on scaling analysis, we develop the contact time model that successfully predicts the contact time measured under a wide range of experiment conditions such as different impact velocities and curvature diameters. We believe that our results illuminate the underlying mechanism for the asymmetric spreading over the curved surface, while the proposed contact time model can be utilized for the design of superhydrophobic surfaces for various thermal applications, where the thermal exchange between the surface and the water drop occurs via a direct physical contact.

11.
Lab Chip ; 20(9): 1601-1611, 2020 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-32249884

RESUMO

Regulating the fluid flow in microfluidic devices enables a wide range of assay protocols for analytical applications. A programmable, photo-paper-based microfluidic device fabricated by using a method of cutting and laminating, followed by printing, is reported. The flow distance of fluid in the photo-paper-based channel was linearly proportional to time. By printing silver nanoparticle (AgNP) and poly[4,5-difluoro-2,2-bis(trifluoromethyl)-1,3-dioxole-co-tetrafluoroethylene] (PTFE) patterns on the surface of a photo-paper-based channel, we were able to either increase or decrease the fluid flow in the fabricated microfluidic devices, while maintaining the linearity in the flow distance-time relation. In comparison to the speed of fluid flow in a pristine channel, by using hydrophilic AgNP patterns, we were able to increase the speed in the channel by up to 15 times while we were able to slow the speed by a factor of 3 when using hydrophobic PTFE dots. We then further demonstrated a single-step protocol for detecting glucose and a multi-step protocol for detecting methyl paraoxon (MPO) with our methods in photo-paper-based microfluidic devices. This approach can lead to improved fluid handling techniques to achieve a wide range of complex, but programmable, assays without the need for any additional auxiliary devices for automated operation.

12.
J R Soc Interface ; 16(161): 20190556, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31795862

RESUMO

Shaping a plant root into an ideal structure for water capture is increasingly important for sustainable agriculture in the era of global climate change. Although the current genetic engineering of crops favours deep-reaching roots, here we show that nature has apparently adopted a different strategy of shaping roots. We construct a mathematical model for optimal root length distribution by considering that plants seek maximal water uptake at the metabolic expenses of root growth. Our theory finds a logarithmic decrease of root length density with depth to be most beneficial for efficient water uptake, which is supported by biological data as well as our experiments using root-mimicking network systems. Our study provides a tool to gauge the relative performance of root networks in transgenic plants engineered to endure a water deficit. Moreover, we lay a fundamental framework for mechanical understanding and design of water-absorptive growing networks, such as medical and industrial fluid transport systems and soft robots, which grow in porous media including soils and biotissues.


Assuntos
Modelos Biológicos , Raízes de Plantas/crescimento & desenvolvimento , Solo , Água/metabolismo
13.
Lab Chip ; 19(10): 1790-1796, 2019 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-30964487

RESUMO

We propose a portable, non-powered, long-term working suction pump with a high flow rate for microfluidic devices. The pump is driven by a superabsorbent polymer enclosed in a housing with porous fins to accelerate water absorption. We experimentally demonstrate that the pump creates an outstanding flow rate of more than 80 µl min-1 and an absorption volume of ∼20 ml. We address the key design principles underlying the outstanding performance of the pump. As an exemplary application, we constructed a portable power generator by combining the hydrogel pump with a reverse electrodialysis (RED) device. This portable system, powered only by KCl solutions with different ion concentrations, exhibited an output density of ∼70 µW cm-2 for more than an hour. The proposed versatile hydrogel pump could provide a breakthrough for developing various portable microfluidic systems.

14.
PLoS One ; 14(1): e0204191, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30703086

RESUMO

In the airway network of a human lung, the airway diameter gradually decreases through multiple branching. The diameter reduction ratio of the conducting airways that transport gases without gas exchange is 0.79, but this reduction ratio changes to 0.94 in acinar airways beyond transitional bronchioles. While the reduction in the conducting airways was previously rationalized on the basis of Murray's law, our understanding of the design principle behind the acinar airways has been far from clear. Here we elucidate that the change in gas transfer mode is responsible for the transition in the diameter reduction ratio. The oxygen transfer rate per unit surface area is maximized at the observed geometry of acinar airways, which suggests the minimum cost for the construction and maintenance of the acinar airways. The results revitalize and extend the framework of Murray's law over an entire human lung.


Assuntos
Bronquíolos/anatomia & histologia , Modelos Biológicos , Oxigênio/metabolismo , Alvéolos Pulmonares/anatomia & histologia , Respiração , Células Acinares/fisiologia , Bronquíolos/citologia , Bronquíolos/fisiologia , Humanos , Tamanho do Órgão/fisiologia , Alvéolos Pulmonares/fisiologia
15.
Int J Oncol ; 53(3): 1269-1278, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29956753

RESUMO

Resveratrol (RSV) is a polyphenolic compound that naturally occurs in grapes, peanuts and berries. Considerable research has been conducted to determine the benefits of RSV against various human cancer types. Tristetraprolin (TTP) is an AU-rich element-binding protein that regulates mRNA stability and has decreased expression in human cancer. The present study investigated the biological effect of RSV on TTP gene regulation in colon cancer cells. RSV inhibited the proliferation and invasion/metastasis of HCT116 and SNU81 colon cancer cells. Furthermore, RSV induced a dose-dependent increase in TTP expression in HCT116 and SNU81 cells. The microarray experiment revealed that RSV significantly increased TTP expression by downregulating E2F transcription factor 1 (E2F1), a downstream target gene of TTP and regulated genes associated with inflammation, cell proliferation, cell death, angiogenesis and metastasis. Although TTP silencing inhibited TTP mRNA expression, the expression was subsequently restored by RSV. Small interfering RNA-induced TTP inhibition attenuated the effects of RSV on cell growth. In addition, RSV induced the mRNA-decaying activity of TTP and inhibited the relative luciferase activity of baculoviral IAP repeat containing 3 (cIAP2), large tumor suppressor kinase 2 (LATS2), E2F1, and lin­28 homolog A (Lin28) in HCT116 and SNU81 cells. Therefore, RSV enhanced the inhibitory activity of TTP in HCT116 and SNU81 cells by negatively regulating cIAP2, E2F1, LATS2, and Lin28 expression. In conclusion, RSV suppressed the proliferation and invasion/metastasis of colon cancer cells by activating TTP.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Regulação Neoplásica da Expressão Gênica , Estilbenos/farmacologia , Tristetraprolina/metabolismo , Regiões 3' não Traduzidas , Antineoplásicos Fitogênicos/uso terapêutico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Relação Dose-Resposta a Droga , Regulação para Baixo , Fator de Transcrição E2F1/metabolismo , Humanos , Estabilidade de RNA/efeitos dos fármacos , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Resveratrol , Estilbenos/uso terapêutico , Tristetraprolina/genética
16.
J Colloid Interface Sci ; 520: 11-18, 2018 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-29518697

RESUMO

We present a combined experimental and theoretical investigation of dewetting on substrates with parallel microgrooves. A thin, static liquid film has an equilibrium thickness so as to minimize the sum of the surface free energy and the gravitational potential energy. When the thickness of a liquid film is less than the equilibrium thickness, the film seeks the equilibrium through contraction of the wetted area, which is referred to as dewetting. We experimentally observed the dewetting of thin, metastable liquid films on substrates with parallel microgrooves. The experiments revealed that the films retract in the direction along the grooves and leaves liquid residues with various morphologies. We classify the residue morphologies into three modes and elucidate the dependence of the mode selection on the groove geometry and the equilibrium contact angle of the liquid. We also experimentally examined the dynamic motion of the receding contact lines of the dewetting films, and developed a mechanical model for the receding speed. Our results provide a basis for controlling liquid films using microstructures, which is useful for lubricant-impregnated surface production, painting, spray cooling, and surface cleaning.

17.
Anal Chem ; 88(3): 1682-7, 2016 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-26713779

RESUMO

We present a novel paper-based flow fractionation system for preconcentration and field-flow separation. In this passive fluidic device, a straight channel is divided into multiple daughter channels, each of which is connected with an expanded region. The hydrodynamic resistance of the straight channel is predominant compared with those of expanded regions, so we can create steady flows through the straight and daughter channels. While the expanded regions absorb a great amount of water via capillarity, the steady flow continues for 10 min without external pumping devices. By controlling the relative hydrodynamic resistances of the daughter channels, we successfully divide the flow with flow rate ratios of up to 30. Combining this bifurcation system with ion concentration polarization (ICP), we develop a continuous-flow preconcentrator on a paper platform, which can preconcentrate a fluorescent dye up to 33-fold. In addition, we construct a field-flow separation system to divide two different dyes depending on their electric polarities. Our flow fractionation systems on a paper-based platform would make a breakthrough for point-of-care diagnostics with specific functions including preconcentration and separation.

18.
Ultrason Sonochem ; 29: 563-7, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26215790

RESUMO

We present the results of experimental investigation of ultrasonic washing of textiles. The results demonstrate that cavitation bubbles oscillating in acoustic fields are capable of removing soils from textiles. Since the washing performance is mitigated in a large washing bath when using an ultrasonic transducer, we propose a novel washing scheme by combining the ultrasonic vibration with a conventional washing method utilizing kinetic energy of textiles. It is shown that the hybrid washing scheme achieves a markedly enhanced performance up to 15% in comparison with the conventional washing machine. This work can contribute to developing a novel laundry machine with reduced washing time and waste water.

19.
Ann Lab Med ; 35(1): 50-6, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25553280

RESUMO

BACKGROUND: The bacterium Chlamydia trachomatis is one of the leading causes of sexually transmitted diseases worldwide. Since no simple and effective tool exists to diagnose C. trachomatis infections, we evaluated a novel point-of-care (POC) test, aQcare Chlamydia TRF kit, which uses europium-chelated nanoparticles and a time-resolved fluorescence reader. METHODS: The test performance was evaluated by comparing the results obtained using the novel POC testing kit with those obtained using a nucleic acid amplification test (NAAT), using 114 NAAT-positive and 327 NAAT-negative samples. RESULTS: The cut-off value of the novel test was 20.8 with a detection limit of 0.27 ng/mL. No interference or cross-reactivity was observed. Diagnostic accuracy showed an overall sensitivity of 93.0% (106/114), specificity of 96.3% (315/327), positive predictive value (PPV) of 89.8% (106/118), and negative predictive value (NPV) of 97.5% (315/323). The sensitivity of the novel test was much higher than that of currently available POC tests. Furthermore, the relative ease and short turnaround time (30 min) of this assay enables C. trachomatis-infected individuals to be treated without a diagnostic delay. CONCLUSIONS: This simple and novel test is a potential tool to screen a larger population, especially those in areas with limited resources.


Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Európio/química , Nanopartículas Metálicas/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Chlamydia trachomatis/isolamento & purificação , DNA Bacteriano/química , DNA Bacteriano/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sistemas Automatizados de Assistência Junto ao Leito , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Adulto Jovem
20.
Anal Chim Acta ; 853: 541-547, 2015 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-25467501

RESUMO

Considering the fatal human victims and economic loss by the annual epidemic influenza virus, the development of a rapid and convenient genetic analysis methodology is demanding for timely on-site pathogen detection. In this study, we utilized reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for multiplex target gene amplification, and the resultant amplicons were analyzed on the immunochromatographic strip (ICS) for subtyping influenza A virus. Through the optimized primer design, reaction temperature and time, and concentration of enzymes (Bst DNA polymerase and AMV reverse transcriptase) and dNTP, the HA (H1, H3, and H5 gene) and conserved M gene were amplified. The ICS contains two test lines in addition to a control line in order to detect the presence of the HA and M gene, thereby informing us of influenza virus A type as well as its subtype (H1N1, H3N2, and H5N1). The combination of the multiplex RT-LAMP with the ICS could be complete in 40 min and the pathotyping and subtyping of influenza A virus were performed even with 10 copies of viral RNA templates. Moreover, the subtyping of clinical samples, which were obtained from patients infected by influenza A virus was successfully confirmed using the multiplex RT-LAMP and ICS techniques, showing great feasibility of our methodology for real sample analysis with high speed, simplicity and sensitivity.


Assuntos
Cromatografia de Afinidade , Vírus da Influenza A/genética , Técnicas de Amplificação de Ácido Nucleico , RNA Viral/análise , Sequência de Bases , Colorimetria , Eletroforese em Gel de Ágar , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H3N2/genética , Virus da Influenza A Subtipo H5N1/genética , Vírus da Influenza A/isolamento & purificação , Influenza Humana/microbiologia , Dados de Sequência Molecular
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