Use of a commercial enzyme immunoassay to monitor dengue virus replication in cultured cells.
Current methods for dengue virus quantitation are either time consuming, technically demanding or costly. As an alternative, the commercial enzyme immunoassay Platelia Dengue NS1 AG (Bio-Rad Laboratories) was used to monitor semiquantitatively dengue virus replication in cultured cells. The presence of NS1 protein was evaluated in supernatants from Vero and C6/36 HT cells infected with dengue virus. The amount of NS1 detected in the supernatants of infected cells was proportional to the initial MOI used and to the time of post infection harvest. This immunoassay was also able to detect the presence of NS1 in the supernatants of infected human macrophages. Inhibition of dengue virus replication in C6/36 HT cells treated with lysosomotropic drugs was readily monitored with the use of this assay. These results suggest that the Platelia Dengue NS1 AG kit can be used as a fast and reliable surrogate method for the relative quantitation of dengue virus replication in cultured cells.
Ludert
Juan E
JE
Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del I.P.N. , Mexico City, Mexico. jludert@cinvestav.mx
Mosso
Clemente
C
Ceballos-Olvera
Ivonne
I
del Angel
Rosa M
RM
D000330:Aedes; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003716:Dengue Virus / Q000187:drug effects / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D008264:Macrophages / Q000378:metabolism / Q000821:virology; D051379:Mice; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000032:analysis; D010948:Viral Plaque Assay; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects
C082515:NS1 protein, dengue-1 virus; D017361:Viral Nonstructural Proteins; D000643:Ammonium Chloride