Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba
The SARS-CoV-2 virus generates the COVID-19 disease, and early diagnosis is very important to prolong patient survival and ensure the safety of the population. The objective of this research was to implement a rapid diagnosis of SARS-CoV-2 based on a viral antigen detection technique by immunofluore...
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| Formato: | Artículo revista |
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Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología
2022
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| Acceso en línea: | https://revistas.unc.edu.ar/index.php/med/article/view/39132 |
| Aporte de: |
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I10-R327-article-39132 |
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ojs |
| institution |
Universidad Nacional de Córdoba |
| institution_str |
I-10 |
| repository_str |
R-327 |
| container_title_str |
Revista de la Facultad de Ciencias Médicas de Córdoba |
| format |
Artículo revista |
| topic |
direct immunofluorescence SARS CoV-2 polyclonal/monoclonal antibodies differential diagnosis inmunofluorescencia directa SARS CoV-2 anticuerpos policlonales/monoclonales diagnóstico diferencial |
| spellingShingle |
direct immunofluorescence SARS CoV-2 polyclonal/monoclonal antibodies differential diagnosis inmunofluorescencia directa SARS CoV-2 anticuerpos policlonales/monoclonales diagnóstico diferencial Bravo, MM Frutos, MC Garay, MP Docena, G Rodríguez Lombardi, G Raskovsky, V Adamo, MP Cámara, A Pérez, IN Pedranti, MS Colazzo Salbetti , MB Alonso, RG Zalazar, JA Bernardi, ME López, P Gruppi, A Kassar, MA Urrúa, S López, A Rumbo, M Hiriart , Y Moreno, LB Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| topic_facet |
direct immunofluorescence SARS CoV-2 polyclonal/monoclonal antibodies differential diagnosis inmunofluorescencia directa SARS CoV-2 anticuerpos policlonales/monoclonales diagnóstico diferencial |
| author |
Bravo, MM Frutos, MC Garay, MP Docena, G Rodríguez Lombardi, G Raskovsky, V Adamo, MP Cámara, A Pérez, IN Pedranti, MS Colazzo Salbetti , MB Alonso, RG Zalazar, JA Bernardi, ME López, P Gruppi, A Kassar, MA Urrúa, S López, A Rumbo, M Hiriart , Y Moreno, LB |
| author_facet |
Bravo, MM Frutos, MC Garay, MP Docena, G Rodríguez Lombardi, G Raskovsky, V Adamo, MP Cámara, A Pérez, IN Pedranti, MS Colazzo Salbetti , MB Alonso, RG Zalazar, JA Bernardi, ME López, P Gruppi, A Kassar, MA Urrúa, S López, A Rumbo, M Hiriart , Y Moreno, LB |
| author_sort |
Bravo, MM |
| title |
Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| title_short |
Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| title_full |
Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| title_fullStr |
Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| title_full_unstemmed |
Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba |
| title_sort |
direct immunofluorescence of sars-cov-2: implementation of a rapid diagnosis from covid19 patient samples from cordoba |
| description |
The SARS-CoV-2 virus generates the COVID-19 disease, and early diagnosis is very important to prolong patient survival and ensure the safety of the population. The objective of this research was to implement a rapid diagnosis of SARS-CoV-2 based on a viral antigen detection technique by immunofluorescence (DFI).
Covid-19(+) samples submitted by collaborating healthcare centers were processed following the recommended safety measures and a total of 165 positive and 59 negative imprints were performed. These imprints were applied to optimize/standardize the DFI following the usual procedure. Primary antibodies: polyclonal serum from convalescent patients; gamma globulins and monoclonal antibodies. Conjugates: goat polyclonal secondary antibodies against mouse and human IgG, and mouse against human IgG labeled with FITC. A total of 33 imprints were used for each primary antibody. Sensitivity, specificity, positive and negative predictive value (PPV) and (NPV) of the technique were determined using the statistical program R-medic.
The results show images with good fluorescence patterns defined for each primary antibody, together with the corresponding controls.The evaluation values of the technique were obtained by performing the analyses in comparison with the RT-PCR technique.The results were similar for post-infection serum and post-vaccinal serum: sensitivity of DFI was 82%, specificity was 64%, PPV was 82% and NPV was 64%; post-infection serum with antibody titer: sensitivity of DFI was 84%, specificity was 57%, PPV was 73% and NPV was 73%; Gamma globulin: DFI sensitivity was 85%, specificity was 67%, PPV was 92% and NPV was 50%; Monoclonal Anti-N and Anti-RBD: DFI sensitivity was 92% , specificity was 78%, PPV was 92% and NPV was 78%. Obtaining good values of sensitivity, specificity, PPV and NPV.
This research reports for the first time in Córdoba, the development of a direct diagnostic method of SARS-CoV-2 by DFI "in house", remarkably relevant to be applied in the initial screening as differential diagnosis when this infection is part of seasonal ARIs, going from pandemic to regional endemic. |
| publisher |
Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología |
| publishDate |
2022 |
| url |
https://revistas.unc.edu.ar/index.php/med/article/view/39132 |
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I10-R327-article-391322024-04-15T16:14:45Z Direct immunofluorescence of SARS-CoV-2: implementation of a rapid diagnosis from Covid19 patient samples from Cordoba Inmunofluorescencia directa de SARS-CoV-2: implementación de un diagnóstico rápido a partir de muestras de pacientes con Covid19 de Córdoba Bravo, MM Frutos, MC Garay, MP Docena, G Rodríguez Lombardi, G Raskovsky, V Adamo, MP Cámara, A Pérez, IN Pedranti, MS Colazzo Salbetti , MB Alonso, RG Zalazar, JA Bernardi, ME López, P Gruppi, A Kassar, MA Urrúa, S López, A Rumbo, M Hiriart , Y Moreno, LB direct immunofluorescence SARS CoV-2 polyclonal/monoclonal antibodies differential diagnosis inmunofluorescencia directa SARS CoV-2 anticuerpos policlonales/monoclonales diagnóstico diferencial The SARS-CoV-2 virus generates the COVID-19 disease, and early diagnosis is very important to prolong patient survival and ensure the safety of the population. The objective of this research was to implement a rapid diagnosis of SARS-CoV-2 based on a viral antigen detection technique by immunofluorescence (DFI). Covid-19(+) samples submitted by collaborating healthcare centers were processed following the recommended safety measures and a total of 165 positive and 59 negative imprints were performed. These imprints were applied to optimize/standardize the DFI following the usual procedure. Primary antibodies: polyclonal serum from convalescent patients; gamma globulins and monoclonal antibodies. Conjugates: goat polyclonal secondary antibodies against mouse and human IgG, and mouse against human IgG labeled with FITC. A total of 33 imprints were used for each primary antibody. Sensitivity, specificity, positive and negative predictive value (PPV) and (NPV) of the technique were determined using the statistical program R-medic. The results show images with good fluorescence patterns defined for each primary antibody, together with the corresponding controls.The evaluation values of the technique were obtained by performing the analyses in comparison with the RT-PCR technique.The results were similar for post-infection serum and post-vaccinal serum: sensitivity of DFI was 82%, specificity was 64%, PPV was 82% and NPV was 64%; post-infection serum with antibody titer: sensitivity of DFI was 84%, specificity was 57%, PPV was 73% and NPV was 73%; Gamma globulin: DFI sensitivity was 85%, specificity was 67%, PPV was 92% and NPV was 50%; Monoclonal Anti-N and Anti-RBD: DFI sensitivity was 92% , specificity was 78%, PPV was 92% and NPV was 78%. Obtaining good values of sensitivity, specificity, PPV and NPV. This research reports for the first time in Córdoba, the development of a direct diagnostic method of SARS-CoV-2 by DFI "in house", remarkably relevant to be applied in the initial screening as differential diagnosis when this infection is part of seasonal ARIs, going from pandemic to regional endemic. El virus SARS-CoV-2 genera la enfermedad COVID-19, siendo el diagnóstico temprano muy importante, para prolongar la supervivencia de los pacientes y garantizar la seguridad de la población. El objetivo de esta investigación fue implementar un diagnóstico rápido de SARS-CoV-2 basado en una técnica detectora de antígeno viral por inmunofluorescencia (IFD). Muestras Covid-19(+) remitidas por los centros asistenciales colaboradores se procesaron siguiendo las medidas de seguridad recomendadas, se realizaron un total de 165 improntas positivas y 59 improntas negativas. Dichas improntas aplicaron para optimizar/estandarizar la IFD siguiendo el procedimiento habitual. Los anticuerpos primarios: sueros policlonales de pacientes convalecientes; gammaglobulinas y anticuerpos monoclonales. Los conjugados: anticuerpos secundarios policlonales de cabra contra IgG de ratón y de origen humano, y de ratón contra IgG humana marcados con FITC. Se utilizaron un total de 33 improntas por cada anticuerpo primario. Se determinaron sensibilidad, especificidad, valor predictivo positivo y negativo (VPP) y (VPN) de la técnica utilizando el programa estadístico R-medic. Los resultados muestran imágenes con buenos patrones de fluorescencia definidos para cada anticuerpo primario, junto a los controles correspondientes. Los valores de evaluación de la técnica se obtuvieron realizando los análisis en comparación con la técnica RT-PCR. Los resultados fueron similares para el Suero post-infección y Suero post-vacunal: sensibilidad de la IFD fue de 82%, especificidad de 64%, VPP fue de 82% y VPN fue de 64%; Suero post-infección con título: sensibilidad de la IFD fue de 84%, especificidad de 57%, VPP fue de 73% y VPN fue de 73%; Gammaglobulina: sensibilidad de la IFD fue de 85%, especificidad de 67%, VPP fue de 92% y VPN fue de 50%; Monoclonales Anti-N y Anti-RBD: sensibilidad de la IFD fue de 92% , especificidad de 78%, VPP fue de 92% y VPN fue de 78%. Obteniendo buenos valores de sensibilidad, especificidad, VPP y VPN. Esta investigación comunica por primera vez en Córdoba, el desarrollo de un método diagnóstico directo de SARS-CoV-2 por IFD “in house”, notablemente relevante para ser aplicado en el tamizaje inicial como diagnóstico diferencial cuando esta infección forme parte de las IRAs estacionales, yendo desde la pandemia a la endemia regional. Universidad Nacional Córdoba. Facultad de Ciencias Médicas. Secretaria de Ciencia y Tecnología 2022-10-26 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion texto texto texto https://revistas.unc.edu.ar/index.php/med/article/view/39132 Revista de la Facultad de Ciencias Médicas de Córdoba.; Vol. 79 No. Suplemento JIC XXIII (2022): Suplemento JIC XXIII Revista de la Facultad de Ciencias Médicas de Córdoba; Vol. 79 Núm. Suplemento JIC XXIII (2022): Suplemento JIC XXIII Revista da Faculdade de Ciências Médicas de Córdoba; v. 79 n. Suplemento JIC XXIII (2022): Suplemento JIC XXIII 1853-0605 0014-6722 http://creativecommons.org/licenses/by-nc/4.0 |