Inactivation of Influenza A virus, Adenovirus, and Cytomegalovirus with PAXgene tissue fixative and formalin.
Formalin fixation is known to inactivate most viruses in a vaccine production context, but nothing is published about virus activity in tissues treated with alternative, non-crosslinking fixatives. We used a model assay based on cell culture to test formalin and PAXgene Tissue fixative for their virus-inactivating abilities. MDCK, A549, and MRC-5 cells were infected with Influenza A virus, Adenovirus, and Cytomegalovirus, respectively. When 75% of the cells showed a cytopathic effect (CPE), the cells were harvested and incubated for 15 min, or 1, 3, 6, or 24 hours, with PBS (positive control), 4% formalin, or PAXgene Tissue Fix. The cells were disrupted and the released virus was used to infect fresh MDCK, A549, and MRC-5 cells cultured on cover slips in 24-well plates. The viral cultures were monitored for CPE and by immunocytochemistry (ICC) to record viral replication and infectivity. Inactivation of Adenovirus by formalin occurred after 3 h, while Influenza A virus as well as Cytomegalovirus were inactivated by formalin after 15 min. All three virus strains were inactivated by PAXgene Tissue fixative after 15 min. We conclude that PAXgene Tissue fixative is at least as effective as formalin in inactivating infectivity of Influenza A virus, Adenovirus, and Cytomegalovirus.
1 Department of Pathology, Josephine Nefkens Institute , Erasmus MC, Rotterdam, Netherlands .
Marcel
M
Kap
Georgina I
GI
Arron
M
M
Loibner
Anja
A
Hausleitner
Gintare
G
Siaulyte
Kurt
K
Zatloukal
Jean-Luc
JL
Murk
Peter
P
Riegman
D000256:Adenoviridae / Q000187:drug effects; D000818:Animals; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects; D004285:Dogs; D005404:Fixatives; D005557:Formaldehyde / Q000494:pharmacology*; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects; D061985:Madin Darby Canine Kidney Cells; D016707:Tissue Fixation / Q000379:methods*; D038301:Virus Inactivation / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects*
D005404:Fixatives; D005557:Formaldehyde