Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 48
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Mil Med ; 2021 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-33428750

RESUMO

INTRODUCTION: Dengue fever, caused by any of the four dengue viruses (DENV1-4), is endemic in more than 100 countries around the world. Each year, up to 400 million people get infected with dengue virus. It is one of the most important arthropod-borne viral diseases. Dengue's global presence poses a medical threat to deploying military personnel and their dependents. An accurate diagnosis followed by attentive supportive care can improve outcomes in patients with severe dengue disease. Dengue diagnostic tests based on PCR and ELISA platforms have been developed and cleared by the U.S. FDA. However, these diagnostic assays are laborious and usually require highly trained personnel and specialized equipment, which presents a significant challenge when conducting operations in austere and resource-constrained areas. InBios International, Inc. (Seattle, WA) has developed two rapid and instrument-free immunochromatographic test prototype devices (multiplex and traditional formats) for dengue diagnosis. MATERIALS AND METHODS: To determine the performance of the InBios immunochromatographic tests, 183 clinical samples were tested on both prototype devices. Both assays were performed without any instruments and the results were read in 20 minutes. RESULTS: The traditional format had better overall performance (sensitivity: 97.4%; specificity: 90%) than the multiplex format (sensitivity: 86.9%; specificity: 63.3%). The traditional format was superior in serotype-specific detection with 100% overall sensitivity for DENV1, DENV3, and DENV4 and 93.3% sensitivity for DENV2 compared to the multiplex format (91.7%, 78.3%, 83.3%, and 96.3% for DENV1, 2, 3, and 4, respectively). The traditional format was easier to read than the multiplex format. The multiplex format was simpler and faster to set up than the traditional format. CONCLUSIONS: The InBios traditional format had a better overall performance and readability profile than the multiplex format, while the multiplex format was easier to set up. Both formats were highly sensitive and specific, were easy to perform, and did not require sophisticated equipment. They are ideal for use in resource-limited settings where dengue is endemic. Based on our overall assessment, the traditional format should be considered for further development and used in the upcoming multicenter clinical trial toward FDA clearance.

2.
Mil Med ; 185(9-10): e1440-e1446, 2020 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-32789444

RESUMO

INTRODUCTION: Measles, mumps, and rubella (MMR) and varicella (VAR) vaccines are the two vaccines administered in large recruit training sites (RTS) that require a single-use syringe to be prefilled with the diluent (ie water) before vaccine reconstitution. Since there are no preservatives in either MMR or VAR vaccines, it is critical to maintain the diluent sterile to ensure the sterility of the reconstituted vaccine. The Department of Defense/Defense Health Agency has instructions on reconstitution of lyophilized vaccines and guidelines for their storage. Vaccine manufacturers provide instructions on how to properly store the diluent. However, there is no clear guidance or standard operating procedures regarding the best practice for preparation and storage of the syringes prefilled with diluent. Various RTS across all four services have their respective routines to best fit their vaccination requirements. Currently, there are no available data on the sterility status of the diluent prepared using these various routines before they are used to reconstitute vaccines. MATERIALS AND METHODS: We investigated the sterility of the diluent (ie water) in prefilled syringes prepared using routines practiced at various RTS. Diluent was drawn up into single syringes and was kept under various conditions (4 °C or room temperature for overnight up to 24 hours) used by various RTS. At indicated time, diluent was injected into sterile vials and the sterility of the diluent was determined by monitoring the presence/growth of bacteria (including aerobic bacteria, mycoplasma, and an obligate intracellular bacterium, Coxiella burnetii), fungi, and viruses for up to 21 days after inoculation into proper and specific culture media. Both traditional cell culture and molecular assays were used to demonstrate the presence or absence of contamination that may compromise the sterility of the diluent. RESULTS: Our results demonstrate that the diluent, after being drawn up to fill the syringe, maintains sterility after storage for overnight up to 24 hours at room temperature or 4 °C with or without recapping the syringes, suggesting that current vaccine reconstitution routines practiced at large military RTS are safe. CONCLUSIONS: Our results demonstrate that in spite of variations in current practices used in various RTS, the diluent in the prefilled syringe tested from each site maintains its sterility and was determined to be safe for use in military health system-wide vaccine reconstitution.

3.
Environ Technol ; : 1-10, 2020 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-32741260

RESUMO

The adsorption of copper ions on magnetic adsorbents with amino-functionalized polymers (DETA/SiO2/Fe3O4) has been investigated. In addition, the structure and characterization of magnetic nano-adsorbents were confirmed using FTIR, TGA, TEM, VSM, and BET. The results showed that DETA/SiO2/Fe3O4 exhibited a paramagnetic behaviour and could be easily recovered and quickly separated from a suspension solution. The optimum pH level for adsorption of copper ions was proposed to be in the range of pH level from 5 to 6, and the maximum adsorption capacity (qm ) and activated energy (Ea ) at 298 K was 13.459 mg g-1 and 22.421 kJ mol-1, respectively. The copper ions adsorption behaviour by DETA/SiO2/Fe3O4 was in good agreement with the Langmuir isotherm rather than Freundlich equation. A pseudo-second-order model could best describe the adsorption kinetics, and the optimal condition of copper ions desorption (99%) from DETA/SiO2/Fe3O4 was provided by 1.0 M HNO3. The novel magnetic nano-adsorbents could not only be regenerated by HNO3 solutions but also reused by the recovery of magnetic force without requiring further downstream treatment.

4.
Polymers (Basel) ; 12(7)2020 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-32605115

RESUMO

In this study, composite nanofibers (SF/PCL/CS) for the application of dressings were prepared with silk fibroin (SF), polycaprolactone (PCL), and chitosan (CS) by electrospinning techniques, and the effect of the fiber diameter was investigated using the three-stage Taguchi experimental design method (L9). Nanofibrous scaffolds were characterized by the combined techniques of scanning electron microscopy (SEM) and transmission electron microscopy (TEM), a cytotoxicity test, proliferation tests, the antimicrobial activity, and the equilibrium water content. A signal-to-noise ratio (S/N) analysis indicated that the contribution followed the order of SF to PCL > flow rate > applied voltage > CS addition, possibly owing to the viscosity and formation of the beaded fiber. The optimum combination for obtaining the smallest fiber diameter (170 nm) with a smooth and uniform distribution was determined to be a ratio of SF to PCL of 1:2, a flow rate of 0.3 mL/hr, and an applied voltage of 25 kV at a needle tip-to-collector distance of 15 cm (position). The viability of these mouse fibroblast L929 cell cultures exceeded 50% within 24 hours, therefore SF/PCL/CS could be considered non-toxic according to the standards. The results proposed that the hydrophilic structure of SF/PCL/CS not only revealed a highly interconnected porous construction but also that it could help cells promote the exchange of nutrients and oxygen. The SF/PCL/CS scaffold showed a high interconnectivity between pores and porosity and water uptake abilities able to provide good conditions for cell infiltration and proliferation. The results from this study suggested that SF/PCL/CS could be suitable for skin tissue engineering.

5.
J Nanosci Nanotechnol ; 20(5): 3252-3257, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31635672

RESUMO

This study focused on an iron phthalocyanine compound with aligned CNTs on the surface of a carbon felt electrode (FePc/CNT/C) to enhance the bio-electro-Fenton microbial fuel cell system cathodes reaction rate of hydrogen peroxide and the electrical plate. Experiments of polarization curves and power density, decolorization of Reactive Black 5 (RB5), and scanning electron microscopy (SEM) measured the characteristics of the cathode plate. FePc/CNT/C presented better electrical properties (open-circuit voltage, maximum current density, and maximum power density) than that of CNT/C and C, as FePc is a catalyst and its planar structure could easily adhere to CNT to enhance the reduction reaction at the cathode and provide higher specific surface area. The optimal decolorization of RB5 dye, as achieved with the FePc/CNT/C electrode, was 61.79% among the three cathode electrodes in the bio-electro-Fenton microbial fuel cell system, and the maximum number of hydroxyl radicals was generated for the cathode electrode of FePc/CNT/C. These results suggest that the bio-electro-Fenton microbial fuel cell system could be applied as an energy-saving and efficient approach for dye-containing wastewater treatment.

6.
Mil Med ; 185(Suppl 1): 624-627, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-31498402

RESUMO

INTRODUCTION: Leptospirosis is a global zoonotic disease spread through contact with contaminated water/soil. The US soldiers at the military bases in these countries are extremely vulnerable, as most of them are immunologically naïve to the responsible pathogen. No recent sero-epidemiological data of leptospirosis among US Marines stationed in Japan were available. MATERIALS AND METHODS: In this study, we analyzed the presence of leptospirosis in US Marines stationed in Japan. One thousand posttour sera samples were analyzed by enzyme-linked immunosorbent assay for Leptospira-specific Immunoglobulin G. RESULTS: Among these 1,000 posttour samples, 85 of them were positive and corresponding pretour samples were analyzed by enzyme-linked immunosorbent assay also. Seroconversion occurred for 35 (3.5%) Marines during their assignment to Japan. These results also indicate that 50 Marine personnels were exposed to leptospires before their assignment to Japan, perhaps because of previous exposure to leptospires at home. CONCLUSION: The 5% rate of seroconversion in 2013 and 2014 suggests that leptospirosis is a potential threat for Marines in the endemic region in Japan.

7.
Environ Sci Pollut Res Int ; 26(28): 29435-29448, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31399835

RESUMO

With the rapid development of economy and society, the quality of groundwater is deteriorating under the dual influence of natural factors and anthropogenic factors, and it seriously threatens the safety of human drinking water. Identifying and analyzing the impact of anthropogenic activities is the key to solving this problem. Based on the groundwater problem in Changle County, Shandong Province, P.R. China, 43 groundwater samples were taken and tested. The statistical characteristics of the monitoring data, the groundwater chemical types, spatial distribution of groundwater, and influencing factors were analyzed by using enrichment factor, Mahalanobis distance, grey water footprint, and so on. The analysis results show that the overall water quality of Changle County is poor, and the main over-standard ions are Cl- , SO2- 4, and NO- 3. There is obvious spatial heterogeneity in the groundwater quality. The spatial variation of NO- 3 is affected by structural factors (topography, hydrology, etc.) and random factors (industrial, agricultural, etc.), and the spatial variability of NO- 3 is the most significant. Other water quality indicators are mainly affected by structural factors. The mass concentration of most ions decreases gradually from north to south, and the overall water quality in the southern region is better than that in the northern region. Thus, the supervision of chemical fertilizers and pesticides should be strengthened. The advanced treatment and reuse of wastewater from industrial parks should be promoted to improve the quality of groundwater and ensure the safety of human drinking water.


Assuntos
Fertilizantes/análise , Água Subterrânea/análise , Praguicidas/análise , Poluentes Químicos da Água/análise , Agricultura , China , Água Subterrânea/química , Humanos , Hidrologia , Praguicidas/química , Qualidade da Água
8.
Anal Chem ; 91(15): 9424-9429, 2019 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-31313917

RESUMO

Single-domain antibodies (sdAb), recombinantly produced variable heavy domains derived from the unconventional heavy chain antibodies found in camelids, provide stable, well-expressed binding elements with excellent affinity that can be tailored for specific applications through protein engineering. Complex matrices, such as plasma and serum, can dramatically reduce assay sensitivity. Thus, to achieve highly sensitive detection in complex matrices a highly efficient assay is essential. We produced sdAb as genetically linked dimers, and trimers, each including SpyTag at their C-terminus. The constructs were immobilized onto dyed magnetic microspheres to which SpyCatcher had been coupled and characterized in terms of their performance as capture reagents in sandwich assays. Initial tests on the ability of oriented monomer, dimer, and trimer captures to improve detection versus unoriented constructs in an assay for staphylococcal enterotoxin B spiked into buffer showed the oriented dimer format provided the best sensitivity while offering robust protein production. Thus, this format was utilized to improve a sdAb-based assay for the detection of dengue virus (DENV) nonstructural protein 1 (NS1) in serum. Detection of NS1 from each of the four DENV serotypes spiked into 50% normal human serum was increased by at least a factor of 5 when using the oriented dimer capture. We then demonstrated the potential of using the oriented dimer capture to improve detection of NS1 in clinical samples. This general method should enhance the utility of sdAb incorporated into any diagnostic assay, including those for high consequence pathogens.


Assuntos
Anticorpos Imobilizados/imunologia , Imunoensaio/métodos , Orientação Espacial , Peptídeos/química , Anticorpos de Domínio Único/imunologia , Imunoensaio/normas , Limite de Detecção , Microesferas , Multimerização Proteica , Proteínas não Estruturais Virais/sangue
9.
Mil Med ; 184(7-8): e275-e280, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30690497

RESUMO

INTRODUCTION: US Military and civilian personnel regularly deploy to regions that are endemic for the Hepatitis B virus (HBV), including the Western Pacific, Africa, Eastern Mediterranean, Southeast Asia, and Europe. When patients have life-threatening injuries that require any blood component that is not immediately available, they are typically transfused with locally collected fresh whole blood from a walking blood bank. Currently, there is no simple and easy method for sensitively screening fresh blood in deployed theaters of conflict. MATERIALS AND METHODS: In order to fill the gap, we have developed a loop-mediated isothermal amplification (LAMP) assay to detect the presence of HBV in blood products. The primers were designed to target the gene of the pre-Surface/Surface antigen region of HBV. The amplification reaction mixture was incubated at 60°C for 60 min. The amplicon can be detected by a handheld fluorescence tube scanner or an immune-chromatography test strip. RESULTS: We were able to detect down to 10 copies of viral DNA by LAMP reaction for HBV DNA extracted from HBV-positive plasma. We also identified the optimal heat treatment condition (125°C for 10 min) for plasma specimens without requiring DNA extraction for the LAMP assay. The sensitivity of the assay was evaluated with polymerase chain reaction (PCR) confirmed HBV-positive samples. Using LAMP, we detected HBV in 107 out of 127 (84%) samples. CONCLUSION: This LAMP assay has the potential to be used in resource-limited settings to improve the safety of locally collected blood in endemic regions.


Assuntos
Hepatite B/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Vírus da Hepatite B , Humanos , Técnicas de Diagnóstico Molecular/normas , Técnicas de Diagnóstico Molecular/estatística & dados numéricos , Técnicas de Amplificação de Ácido Nucleico/normas , Técnicas de Amplificação de Ácido Nucleico/estatística & dados numéricos , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/estatística & dados numéricos , Sensibilidade e Especificidade
11.
Am J Trop Med Hyg ; 99(2): 266-274, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29943710

RESUMO

Leptospirosis is a neglected zoonotic disease with worldwide endemicity and continues to be a significant public health burden on resource-limited populations. Previously, we produced three highly purified recombinant antigens (rLipL32, rLipL41, and rLigA-Rep) and evaluated their performance of detecting Leptospira-specific antibodies in enzyme-linked immunosorbent assay (ELISA) as compared with the microscopic agglutination test (MAT). The overall sensitivity of this assay approached 90%. Recently, another recombinant antigen (rLigB-Rep) was prepared. We tested each individual antigen and a 1:1:1:1 mixture of these four antigens for the detection of Leptospira-specific antibodies in ELISA. The performance of these recombinant antigens was evaluated with a much larger febrile patient panel (337 MAT-confirmed positive sera and 92 MAT-negative sera from febrile patients). Combining the detection results of immunoglobulin M and immunoglobulin G from these four individual antigens, the overall sensitivity was close to 90% but the specificity was only 66%, based on the MAT reference method. The overall sensitivity and specificity of the four-antigen mixture were 82% and 86%, respectively. The mixture of four antigens also exhibited a broader reactivity with MAT-positive samples of 18 serovars from six major pathogenic Leptospira species. Given the limitations of MAT, the data were further analyzed by Bayesian latent class model, showing that ELISA using a 1:1:1:1 mixture still maintained high sensitivity (79%) and specificity (88%) as compared with the sensitivity (90%) and specificity (83%) of MAT. Therefore, ELISA using a mixture of these four antigens could be a very useful test for seroprevalence studies.


Assuntos
Anticorpos Antibacterianos/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Leptospira/imunologia , Leptospirose/diagnóstico , Testes Sorológicos/métodos , Zoonoses/diagnóstico , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Teorema de Bayes , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Leptospirose/imunologia , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade , Zoonoses/imunologia
12.
J Nanosci Nanotechnol ; 18(3): 2241-2248, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29448754

RESUMO

Recyclable magnetites with thioureido group (poly-allyl-thiourea/oleic acid/magnetite, PAT-adsorbent) and amine functional group (ethylenediamine/methyl methacrylate/oleic acid/magnetite, EDA-adsorbent) were synthesized by modifying magnetite with oleic acid, methyl methacrylate, allyl thiourea and ethylenediamine. PAT-adsorbent and EDA-adsorbent were used and compared for adsorption of copper ions in a batch system due to the existence of amino group (-NH2) both on thioureido group and amine functional group. The kinetics of both PAT-adsorbent and EDA-adsorbent were evaluated utilizing pseudo-first-order and pseudo-second-order kinetic models. The equilibrium data was analyzed and compared using the Langmuir and Freundlich isotherm models. The maximum adsorption capacity (Qm) of PAT-adsorbent (19.126 mg g-1) was higher than that of EDA-adsorbent (7.096 mg g-1). As compared to EDA-adsorbent the magnetic adsorbent (PAT-adsorbent) with good desorption performance (>85% desorption efficiency) and easily reuse (>85% recovery by magnetic force) was the important factors for its potential practical application.

13.
Environ Pollut ; 233: 109-113, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29059625

RESUMO

Recent studies suggest that exposure to air pollution might be associated with severity of sleep-disordered breathing (SDB). However, the association between air pollution exposure, especially particulate matter with aerodynamic diameters <= 2.5 µm (PM2.5), and SDB is still unclear. We collected 4312 participants' data from the Taipei Medical University Hospital's Sleep Center and air pollution data from the Taiwan Environmental Protection Administration. Associations of particulate matter with aerodynamic diameters <=10 µm (PM10), PM2.5, nitrogen dioxide (NO2), ozone (O3) and sulfur dioxide (SO2) with apnea-hypopnea index (AHI) and oxygen desaturation index (ODI) were investigated by generalized additive models. We found that an interquartile range (IQR) increase in 1-year mean PM2.5 (3.4 µg/m3) and NO2 (2.7 ppb) was associated with a 4.7% and 3.6% increase in AHI, respectively. We also observed the association of an IQR increase in 1-year mean PM2.5 with a 2.5% increase in ODI. The similar pattern was found in the association of daily mean PM2.5 exposure with increased AHI. Moreover, participants showed significant AHI and ODI responses to air pollution levels in spring and winter. We concluded that exposure to PM2.5 was associated with SDB. Effects of air pollution on AHI and ODI were significant in spring and winter.


Assuntos
Poluentes Atmosféricos/análise , Poluição do Ar/estatística & dados numéricos , Exposição Ambiental/estatística & dados numéricos , Material Particulado/análise , Síndromes da Apneia do Sono/epidemiologia , Adulto , Poluição do Ar/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dióxido de Nitrogênio/análise , Ozônio/análise , Projetos de Pesquisa , Estações do Ano , Dióxido de Enxofre , Taiwan/epidemiologia
14.
Heliyon ; 3(10): e00415, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29057336

RESUMO

Coxiella burnetii, the causative pathogen for Q fever, is an obligate intracellular bacterium and designated as a biosafety level 3 agent. Detection and quantification of the bacteria with conventional culturing methods is time-consuming and poses significant health risks. Polymerase chain reaction (PCR)-based assays have been developed for detecting C. burnetii and could provide rapid diagnosis. However, they require specialized equipment, including a cold chain for PCR reagents that maintains their stability during storage and transport. These requirements limit the advantage of PCR-based methods, especially in resource-limited areas. Previously, we had developed a lyophilized loop-mediated isothermal amplification (LAMP) assay to detect the presence of C. burnetii. To simplify and improve this assay, the reagents for the LAMP assay and the detecting reagent, SYBR green, were lyophilized together. The stability of the lyophilized reagents was evaluated by measuring changes in detection limit for plasmid DNA encoding a C. burnetii gene upon storage at 4 °C, 25 °C, or 37 °C. Our data indicate that the lyophilized reagents remain stable for 24 months when stored at 4 °C, 28 days at 25 °C, and 2 days at 37 °C. This improved LAMP assay can be easily performed in a simple water bath or heating block. The stability at ambient temperature, the simplicity of assay procedure, and the availability of low cost equipment make this method ideal for use in resource-limited settings where Q fever is endemic.

15.
Mil Med ; 182(S1): 369-375, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28291501

RESUMO

Scrub typhus (ST) is an infection caused by Orientia tsutsugamushi. Historically, ST was ranked as the second most important arthropod-borne medical problem only behind malaria during World War II and the Vietnam War. The disease occurs mainly in Southeast Asia and has been shown to emerge and reemerge in new areas, implying the increased risk for U.S. military and civilian personnel deployed to these regions. ST can effectively be treated by doxycycline provided the diagnosis is made early, before the development of severe complications. Scrub Typhus Detect is a lateral flow rapid test based on a mixture of recombinant 56-kDa antigens with broad reactivity. The performance of this prototype product was evaluated against indirect immunofluorescence assay, the serological gold standard. Using 249 prospectively collected samples from Thailand, the sensitivity and specificity for IgM was found to be 100% and 92%, respectively, suggesting a high potential of this product for clinical use. This product will provide a user friendly, rapid, and accurate diagnosis of ST for clinicians to provide timely and accurate treatments of deployed personnel.


Assuntos
Testes Diagnósticos de Rotina/normas , Tifo por Ácaros/diagnóstico , Ásia Sudeste , Testes Diagnósticos de Rotina/instrumentação , Testes Diagnósticos de Rotina/métodos , Diagnóstico Precoce , Febre/etiologia , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Militares , Orientia tsutsugamushi/patogenicidade , Tifo por Ácaros/complicações , Tifo por Ácaros/fisiopatologia , Sensibilidade e Especificidade
16.
J Glob Infect Dis ; 9(1): 3-7, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28250618

RESUMO

BACKGROUND: Leptospirosis is a zoonotic disease which requires laboratory diagnosis for confirmation. MATERIALS AND METHODS: In this study serum samples from adults with acute undifferentiated fever (duration ≤15 days) were tested for IgM antibodies to Leptospira by ELISA, PCR for rrs gene and loop-mediated isothermal amplification (LAMP) assay for LipL32 and LipL41. RESULTS: Among the 150 sera tested, three were positive by PCR, LAMP and IgM ELISA/modified Faines' criteria, two by only PCR; seven only by LAMP assay and forty fulfilled modified Faine's criteria (illness clinically compatible and IgM ELISA positive for leptospirosis). Clinical correlation revealed renal compromise, low platelet count and severe jaundice were significantly related to leptospirosis (P < 0.05). CONCLUSION: This study suggests that LAMP assay could be useful for diagnosis of leptospirosis during the 1st week of illness whereas IgM ELISA forms the mainstay of diagnosis from the 2nd week onward. Further studies especially community based, comparing ELISA, PCR, LAMP, culture and microscopic agglutination test are required to evaluate the veracity of these findings.

17.
Mil Med ; 181(5 Suppl): 227-31, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27168577

RESUMO

Leptospirosis is considered to be the most widespread zoonosis. This worldwide emerging infectious disease is caused by the pathogenic species belonging to the genus Leptospira. Polymerase chain reaction (PCR)-based diagnostic assays have been developed for detecting Leptospira DNA in cell cultures and clinical samples. Because PCR requires specialized equipment and extensive end-user training, it is not suitable for routine work in resource-limited areas. We have developed a loop-mediated isothermal amplification (LAMP) assay to detect the presence of Leptospira in patient, animal and environmental samples using lyophilized reagents at a single temperature of around 63°C with a heating block. The sensitivity of this LAMP assay is very similar to the PCR method. The amplified DNA products can be visualized with the naked eyes using hydroxy naphthol blue or detected by the fluorescence signal of SYBR green dye in the reaction when an ultraviolet lamp or compact fluorescence tube scanner is used. This LAMP assay is simple, rapid, and can be performed with a water bath or heating block. The lyophilized LAMP reagents are stable for 3 months when stored at 4°C and 1 month when stored at 25°C, respectively. It is ideal for resource-limited settings where leptospirosis is endemic.


Assuntos
Leptospira/genética , Leptospirose/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Humanos , Leptospira/patogenicidade , Leptospirose/genética , Técnicas de Amplificação de Ácido Nucleico/normas , Reação em Cadeia da Polimerase em Tempo Real/tendências , Sensibilidade e Especificidade
18.
J Vis Exp ; (110)2016 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-27167168

RESUMO

Coxiella burnetii, the agent causing Q fever, is an obligate intracellular bacterium. PCR based diagnostic assays have been developed for detecting C. burnetii DNA in cell cultures and clinical samples. PCR requires specialized equipment and extensive end user training, and therefore, it is not suitable for routine work especially in a resource-constrained area. We have developed a loop-mediated isothermal amplification (LAMP) assay to detect the presence of C. burnetii in patient samples. This method is performed at a single temperature around 60 °C in a water bath or heating block. The sensitivity of this LAMP assay is very similar to PCR with a detection limit of about 25 copies per reaction. This report describes the preparation of the reaction using lyophilized reagents and visualization of results using hydroxynaphthol blue (HNB) or a UV lamp with fluorescent intercalating dye in the reaction. The LAMP reagents were lyophilized and stored at room temperature (RT) for one month without loss of detection sensitivity. This LAMP assay is particularly robust because the reaction mixture preparation does not involve complex steps. This method is ideal for use in resource-limited settings where Q fever is endemic.


Assuntos
Coxiella burnetii/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Primers do DNA , Humanos , Indicadores e Reagentes , Sensibilidade e Especificidade
19.
J Nanosci Nanotechnol ; 15(4): 2829-34, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26353500

RESUMO

In this study, TiO2 nanoparticles were synthesized from titanium tetraisopropanol (TTIP) using a microwave plasma torch (MPT) and characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and thermogravimetry analysis (TGA). The visible light photocatalysis was studied by the decomposition of methylene blue. MB present in the aqueous solution could be almost completely (> 70%) decomposed within about 720 min of reaction time under visible light irradiation. This is due to the carbon-compounds on the surface of TiO2 (TiOC) corresponding to the results of FTIR. Furthermore, a decrease in recombination between the electron and hole was induced by the existence of TiOC.

20.
J Nanosci Nanotechnol ; 15(4): 2850-7, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26353504

RESUMO

A magnetic adsorbent, amine-functionalized silica magnetite (NH2-Al/SiO2/Fe3O4), has been synthesized to behave as an cationic adsorbent by adjusting the pH value of the aqueous solution to make amino groups protonated. NH2-Al/SiO2/Fe3O4 was used to adsorb phosphate ions in an aqueous solution in a batch system, and the maximum adsorption were found to occur at pH 3.0. The adsorption equilibrium data were all fitted the Langmuir isotherm equation reasonably well, and the maximum adsorption capacities of phosphate ions were more than 40 mg g(-1) and increased with elevating temperature. The enthalpy (ΔH0) and entropy (ΔS0) values of NH2-Al/SiO2/Fe3O4 with the adsorption reaction of phosphate ions were 11.98 KJ mol(-1) and 0.095 KJ (T mol)(-1), respectively. A pseudo-second-order model also could best describe the adsorption kinetics, and the derived activation energy for phosphate ions was 20.2 kJ mol(-1). The optimum condition to desorb phosphate ions from NH2-Al/SiO2/Fe3O4 is provided by a solution with 0.05 M NaOH.


Assuntos
Óxido Ferroso-Férrico/química , Fosfatos/química , Poliaminas/química , Adsorção , Fosfatos/isolamento & purificação , Dióxido de Silício/química , Termodinâmica , Água
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...