A specific polymerase chain reaction based on the gyrB gene sequence and subsequent restriction fragment length polymorphism analysis of Pasteurella pneumotropica isolates from laboratory mice.
For a molecular identification and typing tool, we developed a specific polymerase chain reaction (PCR) based on the gyrB gene sequence and a subsequent restriction fragment length polymorphism (RFLP) analysis using the products amplified from the specific PCR to facilitate discrimination of biotypes of Pasteurella pneumotropica from laboratory mice. Appropriate PCR products, a 1039-basepair fragment for biotype Jawetz and a 1033-basepair fragment for biotype Heyl, were amplified by use of the primers CZO-1 and NJO-2 from all 105 P. pneumotropica isolates tested and the 2 reference strains but not from other bacterial species tested. MspI digestion of PCR-generated products showed 3 RFLP patterns among the 105 isolates, and these patterns correlated with the biotype of the isolate (RFLP pattern 1, biotype Jawetz; RFLP pattern 2, biotype Heyl; and RFLP pattern 3, biotype Jawetz with Beta-hemolytic activity). Our procedure identifies and biotypes isolates of P. pneumotropica from laboratory mice, using simple PCR and enzymatic restriction techniques. Therefore, this procedure is useful as a rapid identification and biotyping tool for isolates of P. pneumotropica from laboratory mice.
Hayashimoto
Nobuhito
N
ICLAS Monitoring Center, Central Institute for Experimental Animals, Kanagawa, Japan. nobuh@ciea.or.jp
Ueno
Masami
M
Takakura
Akira
A
Itoh
Toshio
T
D000818:Animals; D015373:Bacterial Typing Techniques; D027081:DNA Gyrase / Q000737:chemistry / Q000235:genetics*; D017931:DNA Primers; D004262:DNA Restriction Enzymes; D051379:Mice; D008815:Mice, Inbred Strains / Q000382:microbiology*; D008969:Molecular Sequence Data; D044142:Pasteurella pneumotropica / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics
D017931:DNA Primers; D012336:RNA, Ribosomal, 16S; D004262:DNA Restriction Enzymes; D027081:DNA Gyrase