Viral culture and immunofluorescence for the detection of SARS-CoV-2 infectivity in RT-PCR positive respiratory samples

Background: Knowing how long SARS-CoV-2-positive individuals can remain infective is crucial for the design of infection prevention and control strategies. Viral culture is the gold standard for detecting an active-replicative virus and evaluating its infectious potential. Objective: To assess the c...

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Detalles Bibliográficos
Autores: Berengua, Carla|||0000-0001-9963-3809, López Ruiz, Marina, Esteban, Montserrat, Marín, Pilar, Ramos, Paula, del Cuerpo Casas, Margarita|||0000-0001-7973-5496, Gich, Ignasi|||0000-0003-3975-6588, Navarro Risueño, Ferran|||0000-0002-4302-2838, Miró, Elisenda|||0000-0003-2781-6926, Rabella, Núria|||0000-0002-3409-0468
Tipo de recurso: artículo
Fecha de publicación:2022
País:España
Institución:Universitat Autònoma de Barcelona
Repositorio:Dipòsit Digital de Documents de la UAB
Idioma:inglés
OAI Identifier:oai:ddd.uab.cat:285160
Acceso en línea:https://ddd.uab.cat/record/285160
https://dx.doi.org/urn:doi:10.1016/j.jcv.2022.105167
Access Level:acceso abierto
Palabra clave:COVID-19
Infectivity
SARS-CoV-2
VERO E6 cells
Viral culture
Viral isolation
SDG 3 - Good Health and Well-being
Descripción
Sumario:Background: Knowing how long SARS-CoV-2-positive individuals can remain infective is crucial for the design of infection prevention and control strategies. Viral culture is the gold standard for detecting an active-replicative virus and evaluating its infectious potential. Objective: To assess the correlation of SARS-CoV-2 infectivity with the number of days from symptom onset and the Ct value, using culture as a reference method. Also, to describe a detailed protocol for SARS-CoV-2 culture and immunofluorescence confirmation based on our experience with other respiratory viruses. Study design: 100 consecutive respiratory samples positive for SARS-CoV-2 by RT-PCR from different subjects were inoculated into VERO E6 cells. Results: Viral isolation was successful in 58% of samples. The median number of days from symptom onset for culture-positive samples was 2, and 15 for culture-negative samples. Six positive cultures were obtained in patients ≥14 days after symptom onset, all of whom were immunocompromised orwith severe COVID-19. The mean Ct value was 12.64 units higher in culture-negative than in culture-positive samples. The probability of successfully isolating SARS-CoV-2 in samples with a Ct value <22 was 100%, decreasing to 3.1% when >27. Conclusions: Our findings show a significant positive correlation between the probability of isolating SARS-CoV-2 in culture, fewer days of symptoms and a lower RT-PCR Ct value. SARS-CoV-2 infectivity lasts no more than 14 days from symptom onset in immunocompetent individuals. In contrast, in immunocompromised patients or those with severe COVID-19 infectivity may remain after 14 days. Ct value <22 always indicates infectivity.