Detection of rodent coronaviruses in tissues and cell cultures by using polymerase chain reaction.
A polymerase chain reaction (PCR) method was developed for the detection of rodent coronaviruses in biological material by using reverse transcriptase and two primers which flanked an M gene sequence of 375 bp. PCR detected all of 11 different strains of mouse hepatitis virus (MHV) as well as rat sialodacryoadenitis virus but not bovine coronavirus or human coronavirus strains OC43 and 229E. The M gene sequences of bovine coronavirus and human coronavirus OC43 are homologous to that of MHV, but minor differences exist in the primer regions, preventing annealing of the primers. For detecting MHV-Y in tissue samples, PCR was faster than and at least as sensitive as either of the two bioassays (infant mouse bioassay and mouse antibody production test) currently used for MHV diagnostic purposes.
Homberger
F R
FR
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Smith
A L
AL
Barthold
S W
SW
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D001483:Base Sequence; D001681:Biological Assay; D002460:Cell Line; D002478:Cells, Cultured; D003333:Coronaviridae Infections / Q000175:diagnosis; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D051381:Rats
D000914:Antibodies, Viral