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1.
Microbiol Spectr ; 11(6): e0144623, 2023 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-37811937

RESUMO

IMPORTANCE: In this study, we confirmed the binding of M13KO7 to Potato virus Y (PVY) using enzyme-linked immunosorbent assay. M13KO7 is a "bald" bacteriophage in which no recombinant antibody is displayed. M13KO7 is easy to propagate by using Escherichia coli, making this method more reasonable in economic perspective. Based on this study, we suggest that M13KO7 detection system has applicability as a novel biological tool for the detection of PVY.


Assuntos
Bacteriófagos , Potyvirus , Bacteriófagos/genética , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Doenças das Plantas
2.
Mar Biotechnol (NY) ; 24(5): 979-990, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36071349

RESUMO

Viral hemorrhagic septicemia virus (VHSV), one of the most important viral marine pathogens worldwide, has a broad range of hosts, such as members of the families Salmonidae and Paralichthyidae. In addition to being highly contagious, VHSV causes high lethality. The transmission of VHSV can be both vertical and horizontal. In fish, the resolution of VHSV infection is challenging. Thus, early diagnosis of VHSV infections is critical, especially in fish farms that have a high population of juvenile fish. Serological methods are commonly used to detect viral antigens. However, limited serological methods are available for marine viruses. In this study, a VHSV-specific single-chain variable fragment (scFv), E5, was selected using the yeast surface display and phage display systems. scFv, a type of recombinant antibody, comprises a variable heavy chain ([Formula: see text]) and a variable light chain ([Formula: see text]) connected by a polypeptide linker. An scFv clone was selected from the VHSV glycoprotein-expressing yeast cells using the bio-panning method. The scFv-encoding gene was subcloned and expressed in the Escherichia coli expression system. The binding affinity of the expressed and purified scFv protein was determined using an enzyme-linked immunosorbent assay and western blotting. Thus, this study reported a method to identify VHSV-specific scFv using bio-panning that can be utilized to develop a diagnostic system for other viruses.


Assuntos
Doenças dos Peixes , Septicemia Hemorrágica Viral , Novirhabdovirus , Anticorpos de Cadeia Única , Animais , Antígenos Virais , Doenças dos Peixes/diagnóstico , Glicoproteínas , Septicemia Hemorrágica Viral/diagnóstico , Novirhabdovirus/genética , Saccharomyces cerevisiae , Anticorpos de Cadeia Única/genética
3.
Virusdisease ; 31(3): 251-256, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32904820

RESUMO

Red sea bream iridovirus (RSIV) and Viral hemorrhagic septicemia virus (VHSV) are the most important viral marine pathogens in South Korea because RSIV and VHSV infect and cause high mortality rates in major fish species such as Paralichthys olivaceus and Sebastes schlegelii. These viruses can be transmitted both vertically and horizontally, and early diagnosis is imperative. In this research, RSIV and VHSV viral genomes are detected by PCR-lateral flow assay (LFA). PCR-LFA is sensitive, capable of detecting a viral genome at a concentration of 2-200 fg/µL. Development of this detection method is very meaningful because LFA is simple, requiring a minimum of personnel training to perform. Additionally, LFA requires less time than other detection methods and can be an immediate detection tool that is indispensable in preventing rapid viral spread.

4.
Sci Rep ; 6: 19013, 2016 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-26743765

RESUMO

Tomato yellow leaf curl virus (TYLCV) is one of the most well-known tomato-infecting begomoviruses and transmitted by Bemisia tabaci. Seed transmission has previously been reported for some RNA viruses, but TYLCV has not previously been described as a seed-borne virus. In 2013 and 2014, without whitefly-mediated transmission, TYLCV was detected in young tomato plants germinated from fallen fruits produced from TYLCV-infected tomato plants in the previous cultivation season. In addition, TYLCV-Israel (TYLCV-IL) was also detected in seeds and their seedlings of TYLCV-infected tomato plants that were infected by both viruliferous whitefly-mediated transmission and agro-inoculation. The seed infectivity was 20-100%, respectively, and the average transmission rate to seedlings was also 84.62% and 80.77%, respectively. TYLCV-tolerant tomatoes also produced TYLCV-infected seeds, but the amount of viral genome was less than seen in TYLCV-susceptible tomato plants. When tomato plants germinated from TYLCV-infected seeds, non-viruliferous whiteflies and healthy tomato plants were placed in an insect cage together, TYLCV was detected from whiteflies as well as receiver tomato plants six weeks later. Taken together, TYLCV-IL can be transmitted via seeds, and tomato plants germinated from TYLCV-infected seeds can be an inoculum source of TYLCV. This is the first report about TYLCV seed transmission in tomato.


Assuntos
Begomovirus/patogenicidade , Genoma Viral , Doenças das Plantas/virologia , Plântula/virologia , Sementes/virologia , Solanum lycopersicum/virologia , Sequência de Aminoácidos , Animais , Begomovirus/fisiologia , Transmissão de Doença Infecciosa , Germinação/fisiologia , Hemípteros/virologia , Insetos Vetores/virologia , Solanum lycopersicum/crescimento & desenvolvimento , Plântula/crescimento & desenvolvimento , Sementes/crescimento & desenvolvimento , Alinhamento de Sequência , Virulência
5.
JRSM Short Rep ; 3(7): 46, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22908027

RESUMO

OBJECTIVES: To conduct an exploratory study to learn about the experiences of GPs who have undertaken international work. DESIGN: Cross-sectional survey SETTING: Online survey of UK-based GPs. Members of all UK RCGP faculties were invited to participate by email and the survey was publicised on the RCGP website PARTICIPANTS: All UK-based GPs MAIN OUTCOME MEASURES: Types of UK and international work undertaken, barriers, competencies gained, influence on career and future plans. RESULTS: The study identified 439 respondents, in a variety of GP roles at all career stages, who had undertaken international work in their role as a doctor. GPs are undertaking international work in both high and low/middle-income countries, engaging in a wide range of clinical and non-clinical activities. Respondents reported gaining a range of competencies from international work, which could be transferred back to the UK setting to a variable degree. Commonly cited barriers to international work were having to leave friends and family, and concerns regarding future employment and pension. Most reported that engaging in international work had influenced the direction of their career, with the largest proportion stating that they wish to work predominantly in the UK, with some international work in the future. CONCLUSION: The study highlights the variety of ways in which UK GPs are combining UK general practice and international work, competencies gained with such work, and ability to transfer these back to the UK setting. Historical barriers to international work still exist and future research could further examine the value of such work.

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