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1.
J Microbiol Methods ; 224: 106985, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38960328

RESUMEN

The assessment of ELISA plates coated with phenolic glycolipid-I/PGL-I revealed excellent stability during eight years of storage at room temperature, promoting consistent IgM antibody detection in multibacillary leprosy patients. These stable, standardized plates can significantly contribute to efficient leprosy serology research and support its widespread distribution and use in endemic countries.


Asunto(s)
Anticuerpos Antibacterianos , Antígenos Bacterianos , Ensayo de Inmunoadsorción Enzimática , Glucolípidos , Inmunoglobulina M , Mycobacterium leprae , Inmunoglobulina M/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Mycobacterium leprae/inmunología , Humanos , Anticuerpos Antibacterianos/inmunología , Glucolípidos/inmunología , Antígenos Bacterianos/inmunología , Lepra/diagnóstico , Lepra/microbiología , Lepra/inmunología , Factores de Tiempo
2.
J Virol Methods ; 329: 114969, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38834144

RESUMEN

The recent COVID-19 pandemic disclosed a critical shortage of diagnostic kits worldwide, emphasizing the urgency of utilizing all resources available for the development and production of diagnostic tests. Different heterologous protein expression systems can be employed for antigen production. This study assessed novel SARS-CoV-2 proteins produced by a transient expression system in Nicotiana benthamiana utilizing an infectious clone vector based on pepper ringspot virus (PepRSV). These proteins included the truncated S1-N protein (spike protein N-terminus residues 12-316) and antigen N (nucleocapsid residues 37-402). Two other distinct SARS-CoV-2 antigens expressed in Escherichia coli were evaluated: QCoV9 chimeric antigen protein (spike protein residues 449-711 and nucleocapsid protein residues 160-406) and QCoV7 truncated antigen (nucleocapsid residues 37-402). ELISAs using the four antigens individually and the same panel of samples were performed for the detection of anti-SARS-CoV-2 IgG antibodies. Sensitivity was evaluated using 816 samples from 351 COVID-19 patients hospitalized between 5 and 65 days after symptoms onset; specificity was tested using 195 samples collected before 2018, from domiciliary contacts of leprosy patients. Our findings demonstrated consistent test sensitivity, ranging from 85 % to 88 % with specificity of 97.5 %, regardless of the SARS-CoV2 antigen and the expression system used for production. Our results highlight the potential of plant expression systems as useful alternative platforms to produce recombinant antigens and for the development of diagnostic tests, particularly in resource-constrained settings.


Asunto(s)
Anticuerpos Antivirales , Antígenos Virales , COVID-19 , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Inmunoglobulina G , Nicotiana , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus , Escherichia coli/genética , Escherichia coli/metabolismo , Humanos , Nicotiana/genética , Inmunoglobulina G/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , SARS-CoV-2/inmunología , SARS-CoV-2/genética , Antígenos Virales/inmunología , Antígenos Virales/genética , COVID-19/diagnóstico , COVID-19/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Glicoproteína de la Espiga del Coronavirus/genética , Prueba Serológica para COVID-19/métodos , Sensibilidad y Especificidad , Proteínas de la Nucleocápside de Coronavirus/inmunología , Proteínas de la Nucleocápside de Coronavirus/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Adulto , Persona de Mediana Edad , Fosfoproteínas
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