Human astrovirus isolation and propagation in multiple cell lines

Laboratory adapted human astrovirus serotypes 1 through 7 were tested for g rowth in 15 human, 7 simian, and 10 other non-primate mammalian cell lines. Propagation of all seven serotypes was successful in the human cell lines Caco-2, T84, HT-29, and in the African green monkey kidney cell line MA-104 . Both primary and secondary African green monkey kidney cells were more ef fective than Rhesus monkey kidney cells for cultivation of astrovirus. Exce pt for human foreskin cells, all of the other human and simian cell lines s upported growth of at least one astrovirus serotype. The only non-primate c ell line that permitted sustained passage of astroviruses was the BHK-21 (C 13) cell line for astrovirus serotype 2. Seventeen human stool specimens th at had previously been shown to be astrovirus positive by ELISA were cultur ed in Caco-2, T84, HT-29, SK-CO-I, PLC/PRF/5, MA-104, and VERO cells. Caco- 2 cells (13 isolates), T84 cells (12 isolates) and PLC/PRF/5 cells (12 isol ates) were the cell lines most effective for isolation of human astroviruse s from clinical stool specimens. By immunofluorescent staining of infected cells, culturing of the same 17 specimens in shell vials for 18 h was posit ive for astroviruses in all 17 specimens in Caco-2 cells, 12 in T84 cells, and 7 in PLC/PRF/5 cells. Shell vial assay is suitable as a rapid and sensi tive culture technique for detection of astroviruses in clinical specimens.