Towards dynamic monitoring of cell cultures using high throughput sequencing.
Vaccine
; 37(7): 1001-1005, 2019 02 08.
Article
en En
| MEDLINE
| ID: mdl-30642729
We used a combination of DOP-PCR with high throughput sequencing (HTS) to study infected cell cultures over time to assess the feasibility of using this technique to provide a read-out other than cytopathic effect in cell culture infectivity assays. Because DOP-PCR primers feature a short constant sequence at their 3' terminus, the procedure yields a reproducible representational library of products from a given PCR template, including viral nucleic acids. Using SV40- and MVM-infected cultures harvested at different times, we show that the number of viral matches among DOP-PCR products parallels the quantity of virus as shown by real-time PCR, and further show that HTS analysis of specific DOP-PCR products that increase in quantity over time could be used to identify the infecting virus with a sensitivity similar to that of typical cell-culture assays that rely on cytopathic effect.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Cultivo de Virus
/
Reacción en Cadena de la Polimerasa
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Virus 40 de los Simios
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Secuenciación de Nucleótidos de Alto Rendimiento
/
Virus Diminuto del Ratón
Límite:
Animals
Idioma:
En
Revista:
Vaccine
Año:
2019
Tipo del documento:
Article
Pais de publicación:
Países Bajos