One-step immunochromatographic assay for the detection of Staphylococcus aureus

[[abstract]]An analytical system of immunochromatographic assay based on gold nanoparticles was developed to detect Staphylococcus aureus. The assay was in the sandwich format, using anti-protein A IgG with two distinct specificities. One anti-protein A IgG was immobilized in a defined detection zone on a porous nitrocellulose membrane, while the other anti-protein A IgG was conjugated with gold nanoparticles. The mixture was then passed along the porous membrane by capillary action past the anti-protein A IgG in the detection zone, binding the particles to which surface protein A was already bound to their surface, yielding a red color. The sensitivity and specificity in the immunochromatographic test were 100 and 93.0–100% for 28 S. aureus strains and 23 non-S. aureus strains, respectively. S. aureus was detected by immunochromatographic testing of two samples of precooked foods naturally contaminated with the bacterium. Eleven processed foods artificially inoculated with S. aureus at <25 CFU/g, all yielded positive results in the immunochromatographic test

Gold nanoparticle-based immunochromatographic Test for Identification of Staphylococcus aureus from Clinical Specimens.

[[abstract]]Background Staphylococcus aureus is one of the most important human pathogens, causing both nosocomial and community-acquired infections. Therefore, a method for rapidly detecting for S. aureus would be useful. We describe an analytical system of immunochromatographic assay based on gold nanoparticles developed for the detection of S. aureus in patient specimen. Methods The assay was in the sandwich format, using anti-protein A IgG with 2 distinct specificities. One anti-protein A IgG was immobilized in a defined detection zone on a porous nitrocellulose membrane, while the other anti-protein A IgG was conjugated with gold nanoparticles. The mixture was then passed along the porous membrane by capillary action past the anti-protein A IgG in the detection zone, binding the particles that to which surface protein A was already bound to their surface, yielding a red color. Results The sensitivity and specificity in the immunochromatographic test were 100% and 94.7–100% for 130 S. aureus strains and 36 non-S. aureus strains, respectively. The results were comparable to the conventional coagulase test and latex agglutination test of different bacteria. Conclusion This method may be useful for analyzing S. aureus in patient specimen. It was quick, easy to perform, and with a long shelf life at room temperature

Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus.

[[abstract]]Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1–10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples

Detection of Escherichia coli using CMOS array photo sensor-based enzyme biochip detection system.

[[abstract]]This work presents optical enzyme detection system based on the CMOS array photo sensor and 1 × 3 polymeric enzyme biochip for detecting Escherichia coli in a one-step procedure. This assay, using 4-methylumbelliferyl-β-d-glucuronide (MUG) as a fluorogenic substrate, had a detection limit of 0.1 U/ml for β-glucuronidase (GUD), which was approximately equal to a cell concentration of 106 CFU/ml of E. coli. MUG was incorporated into lauryl tryptose broth at a final concentration of 100 μg/ml for immediate verification of the presence of E. coli in 1 × 3 polymeric enzyme biochip. The 40 strains of E. coli studied all produced GUD. Of another 36 strains of bacteria tested, one strain (Salmonella choleraesuis subsp. choleraesuis) yielded very small amounts of GUD after 24 h incubation. The optical enzyme detection system was sensitive and rapid

Gold nanoparticle-based immunochromatographic assay for the detection of Staphylococcus aureus.

[[abstract]]An analytical system of immunochromatographic assay based on gold nanoparticles was developed for the detection of Staphylococcus aureus. Staphylococcal protein A, a cell wall protein of S. aureus, has the ability to interact with several host components, possibly indicating a role as a virulence factor in S. aureus infections. The assay was constructed in the form of sandwich by using anti-protein A IgG with two distinct specificities. One anti-protein A IgG was immobilized in a defined detection zone on a porous nitrocellulose membrane, while the other anti-protein A IgG was conjugated with gold nanoparticles. The sample flows along the porous membrane by capillary action past the anti-protein A IgG in the detection zone, binding the particles that to which surface protein A was already bound to their surface, yielding a red color. The rapid observation of results directly by the naked eye ensures the convenience of performing bioassays on field. As the results, the sensitivity in the immunochromatographic test of 306 S. aureus strains were 100% and the specificity of 44 non-S. aureus strains were 96.0 to 100%. Twelve processed food samples inoculated artificially with 0.9, 1.2, 2.4, and 6 CFU/g of S. aureus and all yielded positive results in the immunochromatographic test. As compared to the conventional culture methods which take 5 to 6 days to complete, the immunochromatographic test can detect low numbers of S. aureus in processed foods with a total analytical time of only 25 h

Gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of Japanese encephalitis virus

[[abstract]]Virus isolation and antibody detection are routinely used for diagnosis of Japanese encephalitis virus (JEV) infection, but the low level of transient viremia in some JE patients makes JEV isolation from clinical and surveillance samples very difficult. We describe the use of gold nanoparticle-based RT-PCR and real-time quantitative RT-PCR assays for detection of JEV from its RNA genome. We tested the effect of gold nanoparticles on four different PCR systems, including conventional PCR, reverse-transcription PCR (RT-PCR), and SYBR green real-time PCR and RT-PCR assays for diagnosis in the acute phase of JEV infection. Gold nanoparticles increased the amplification yield of the PCR product and shortened the PCR time compared to the conventional reaction. In addition, nanogold-based real-time RT-PCR showed a linear relationship between Ct and template amount using ten-fold dilutions of JEV. The nanogold-based RT-PCR and real-time quantitative RT-PCR assays were able to detect low levels (1-10 000 copies) of the JEV RNA genomes extracted from culture medium or whole blood, providing early diagnostic tools for the detection of low-level viremia in the acute-phase infection. The assays described here were simple, sensitive, and rapid approaches for detection and quantitation of JEV in tissue cultured samples as well as clinical samples

Detection of Escherichia coli using CMOS array photo sensor-based enzyme biochip detection system

[[abstract]]This work presents optical enzyme detection system based on the CMOS array photo sensor and I x 3 polymeric enzyme biochip for detecting Escherichia coli in a one-step procedure. This assay, using 4-methylumbelliferyl-beta-D-glucuronide (MUG) as a fluorogenic substrate, had a detection limit of 0.1 U/ml for beta-glucuronidase (GUD), which was approximately equal to a cell concentration of 10(6), CFU/ml of E. coli. MUG was incorporated into lauryl tryptose broth at a final concentration of 100 mu g/ml for immediate verification of the presence of E coli in 1 x 3 polymeric enzyme biochip. The 40 strains of E. coli studied all produced GUD. Of another 36 strains of bacteria tested, one strain (Salmonella choleraesuis subsp. choleraesuis) yielded very small amounts of GUD after 24 h incubation. The optical enzyme detection system was sensitive and rapid

Review on the Conflicts between Offshore Wind Power and Fishery Rights: Marine Spatial Planning in Taiwan

In recent years, Taiwan has firmly committed itself to pursue the green energy transition and a nuclear-free homeland by 2025, with an increase in renewable energy from 5% in 2016 to 20% in 2025. Offshore wind power (OWP) has become a sustainable and scalable renewable energy source in Taiwan. Maritime Spatial Planning (MSP) is a fundamental tool to organize the use of the ocean space by different and often conflicting multi-users within ecologically sustainable boundaries in the marine environment. MSP is capable of definitively driving the use of offshore renewable energy. Lessons from Germany and the UK revealed that MSP was crucial to the development of OWP. This paper aims to evaluate how MSP is able to accommodate the exploitation of OWP in Taiwan and contribute to the achievement of marine policy by proposing a set of recommendations. It concludes that MSP is emerging as a solution to be considered by government institutions to optimize the multiple use of the ocean space, reduce conflicts and make use of the environmental and economic synergies generated by the joint deployment of OWP facilities and fishing or aquaculture activities for the conservation and protection of marine environments.Peer Reviewe