Detection of the bunyavirus Germiston in VERO and Aedes albopictus C6/36 cells by in situ hybridization using cDNA and asymmetric RNA probes.
Using Germiston virus infected vertebrate (VERO) and invertebrate (Aedes albopictus C6/36) cells, paraformaldehyde-glutaraldehyde fixative allowed the best preservation of cellular morphology and the highest hybridization signals with cDNA and asymmetric RNA probes against the viral S segment. Asymmetric RNA probes always gave higher sensitivity and better specificity of in situ hybridization than the nick-translated symmetric DNA probe in both vertebrate and invertebrate cells. The study of Aedes albopictus C6/36 cells persistently infected with Germiston virus showed that only a small number of cells contained the S segment, and that the replication and transcription of the S segment took place in the cytoplasm of acutely and persistently infected cells.
Delord
B
B
Laboratoire de Virologie, Université de Bordeaux II, France.
Poveda
J D
JD
Astier-Gin
T
T
Gerbaud
S
S
Fleury
H J
HJ
D000330:Aedes / Q000382:microbiology; D000818:Animals; D002043:Bunyaviridae / Q000302:isolation & purification*; D002460:Cell Line; D015342:DNA Probes; D005404:Fixatives; D015347:RNA Probes; D012367:RNA, Viral / Q000032:analysis*; D014709:Vero Cells
D015342:DNA Probes; D005404:Fixatives; D015347:RNA Probes; D012367:RNA, Viral