TY - JOUR
T1 - Bluetongue virus propagation and plaque assay
T2 - Variation due to medium and serum supplement
AU - Blanchard, Myra
AU - Stott, Jeffrey L
PY - 1989
Y1 - 1989
N2 - Two base media, minimal essential medium (MEM) and RPMI 1640, were supplemented with a variety of serum extenders and/or substitutes for the purpose of defining a medium formulation capable of supporting good cell (VERO) growth and virologic assays. Bluetongue virus (BTV), the prototype Orbivirus in the Reoviridae, was used in all studies. In general, VERO cells grown in RPMI performed better than those grown in MEM relative to cell growth, virus production and plaque assay. RPMI was better for supporting cell growth when serum extenders (NuSerum or SerXtend) were employed as supplements. Relative to virologic techniques, cells grown in RPMI produced higher virus titers in both propagation studies and plaque assays. The interval from infection to >90% cytopathic effect (CPE) was consistently shorter with RPMI as the base medium. Cell cultures supported with RPMI base medium, supplemented with 3.5% FBS with SerXtend, provided the best overall performance relative to: (a) amount of virus produced by infected cell monolayers, (b) sensitivity to productive infection under overlay conditions (revealed the highest titer of a standard virus stock) and (c) plaque assay quality including cell quality, plaque size and plaque clarity.
AB - Two base media, minimal essential medium (MEM) and RPMI 1640, were supplemented with a variety of serum extenders and/or substitutes for the purpose of defining a medium formulation capable of supporting good cell (VERO) growth and virologic assays. Bluetongue virus (BTV), the prototype Orbivirus in the Reoviridae, was used in all studies. In general, VERO cells grown in RPMI performed better than those grown in MEM relative to cell growth, virus production and plaque assay. RPMI was better for supporting cell growth when serum extenders (NuSerum or SerXtend) were employed as supplements. Relative to virologic techniques, cells grown in RPMI produced higher virus titers in both propagation studies and plaque assays. The interval from infection to >90% cytopathic effect (CPE) was consistently shorter with RPMI as the base medium. Cell cultures supported with RPMI base medium, supplemented with 3.5% FBS with SerXtend, provided the best overall performance relative to: (a) amount of virus produced by infected cell monolayers, (b) sensitivity to productive infection under overlay conditions (revealed the highest titer of a standard virus stock) and (c) plaque assay quality including cell quality, plaque size and plaque clarity.
KW - Bluetongue virus
KW - Minimal essential medium
KW - RPMI 1640
KW - Serum extender and supplement
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U2 - 10.1016/0166-0934(89)90038-4
DO - 10.1016/0166-0934(89)90038-4
M3 - Article
C2 - 2547822
AN - SCOPUS:0024384544
VL - 24
SP - 265
EP - 274
JO - Journal of Virological Methods
JF - Journal of Virological Methods
SN - 0166-0934
IS - 3
ER -