Detection of MHV-RNAs in mouse intestines and in filter dust in mouse room ventilation duct by a modified RT-nested PCR.
We applied RT-nested PCR for the detection of MHV genomic RNA in a modified manner to obtain RNA from the intestines of mice and from filter dust in the ventilation ducts of the room in which a contaminated mouse colony was kept. Since the sequences of MHV-RNA that were extracted from the intestine of a serologically MHV-positive mouse in room No. 2 (MS2) and from the filter dust in a ventilation duct in the same room (FD2) were identical, amplified product from filter dust was demonstrated to come from the MHV contaminated room. Furthermore, sequences of FD2 and of filter dust from another contaminated mouse room, No. 7 (FD7) showed 38 nucleotide exchanges among 368-bp (10.3%), suggesting that two different MHV strains were contaminating our facilities. SSCP analysis of Dra I-digested PCR product of 393 bp also showed different patterns in FD2 and FD7 samples.
Oyanagi
Mitsuru
M
Animal Resources Branch, Center for Bioresource-based Research, Brain Researches Institute, Niigata University, Niigata, Japan.
Kato
Akiko
A
Yamada
Yasuko K
YK
Sato
Norimitsu L
NL
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004391:Dust / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D006799:Housing, Animal; D007422:Intestines / Q000821:virology*; D051379:Mice; D008969:Molecular Sequence Data; D006517:Murine hepatitis virus / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004391:Dust; D012367:RNA, Viral