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1.
Int J Biol Macromol ; 253(Pt 3): 126644, 2023 Dec 31.
Artículo en Inglés | MEDLINE | ID: mdl-37659497

RESUMEN

Immunoglobulin Y (IgY) could be used in serological diagnosis focused on several infectious agents. This study aims to produce IgY anti-hepatitis B virus surface antigen (anti-HBs) and to assess its use in enzyme immunoassays. Antibodies were produced by immunizing chickens with Hepatitis B vaccine associated (group A), or not, with adjuvant CpG-ODN (group B). Eggs were collected for 20 weeks, yolks were purified based on using polyethylene glycol and affinity chromatography. IgY anti-HBs was featured based on SDS-PAGE and Western Blot techniques. Total protein concentration was measured through spectrophotometry. In-house ELISA used to detect HBsAg was developed based on using IgG/HRP conjugate and IgY-anti-HBs sensitized microplates. Thus, IgY anti-HBs were confirmed through molecular pattern based on SDS-PAGE, whereas specificity of anti-HBs was confirmed through Western Blot. Mean total protein reached 3.27 ± 3.00 mg/mL and 3.11 ± 3.12 mg/mL in groups A and B, respectively. In-house ELISA was developed based on using a panel of HBV positive and negative serum samples; it recorded 100 % sensitivity and 78.9 % specificity to detect HBsAg. In conclusion, it was possible producing anti-HBs IgY by immunizing chickens with HBV vaccine; this molecule could be used as capture antibody to help detecting HBsAg in-house ELISA.


Asunto(s)
Antígenos de Superficie de la Hepatitis B , Hepatitis B , Animales , Pollos , Vacunas contra Hepatitis B , Hepatitis B/diagnóstico , Antígenos e de la Hepatitis B , Anticuerpos contra la Hepatitis B
2.
Int J Biol Macromol ; 189: 785-791, 2021 Oct 31.
Artículo en Inglés | MEDLINE | ID: mdl-34416265

RESUMEN

Immunoglobulin yolk (IgY) is therapeutic antibodies presented in yolk eggs of birds, reptiles, and amphibians. These proteins produced by the immune system of the animal, are capable of neutralizing antigenic molecules, including viral antigens, fulfilling a role in the body defense. The specificity of these antibodies and the facility for their production, make these molecules capable of being used as tools for diagnosis and immunotherapy. Regarding this last aspect, it is common knowledge that the field of virology, is racing against time in the development of new drugs and vaccines to try to contain pandemics and local epidemics and, in counterproposal, avian antibodies are neutralizing molecules that can help in the control and spread of disease. These molecules have been explored for years and currently chicken eggs are produced in large quantities from the animal's immunization against a specific pathogen. Thus, on this subject, this review made a survey of these researches and presents a summary of all the successful cases and perspectives in the use of IgYs as tools for viral immunization.


Asunto(s)
Antivirales/farmacología , Inmunoglobulinas/farmacología , Animales , Humanos , Inmunización , Inmunoglobulinas/química , Inmunoglobulinas/aislamiento & purificación , Virosis/inmunología , Virosis/terapia , Virus/efectos de los fármacos , Virus/inmunología
3.
BMC Res Notes ; 12(1): 523, 2019 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-31429797

RESUMEN

OBJECTIVE: Dried blood spots (DBSs) can be used as an alternative to serum samples because they are easily collected and can be transported without refrigeration to reference laboratories for diagnosis. The present study was performed to evaluate the utility of electrochemiluminescence immunoassay "ECLIA" for anti-HCV, HBsAg and anti-HBc detection from DBS samples. RESULTS: Anti-HCV was detected in 103 DBS samples from 108 paired, positive serum and undetected in 364 DBS samples from 366 paired, negative specimens, giving a sensitivity of 95.4% and a specificity of 99.4%. HBsAg was detected in 67 DBS samples out of 71 positive, paired serum and was undetected among 295 DBS samples from 298 paired, negative specimens, giving a sensitivity and specificity of 94.4% and 99%, respectively. Anti-HBc was detected in 160 DBS samples from 185 paired, positive serum specimens and undetected in 349 DBS samples from 357 paired, negative serum specimens, giving a sensitivity of 86.5% and a specificity of 97.8%. Overall, the Kappa index indicated a high agreement between results obtained for the serum and DBS samples (k: 0.95, 0.93 and 0.86 for anti-HCV, HBsAg, anti-HBc, respectively). In conclusion, the ECLIA test could be used for detecting hepatitis B and C markers in DBS.


Asunto(s)
Bioensayo/métodos , Biomarcadores/sangre , Pruebas con Sangre Seca/métodos , Hepatitis B/diagnóstico , Hepatitis C/diagnóstico , Automatización , Humanos , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad
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