title: Specific detection and identification of [Actinobacillus] muris by PCR using primers targeting the 16S-23S rRNA internal transcribed spacer regions.
abstract: [Actinobacillus] muris represents along with [Pasteurella] pneumotropica the most prevalent Pasteurellaceae species isolated from the laboratory mouse. Despite the biological and economic importance of Pasteurellaceae in relation to experimental animals, no molecular based methods for the identification of [A.] muris are available. The aim of the present investigation was to develop a PCR method allowing detection and identification of [A.] muris. In this assay, a Pasteurellaceae common forward primer based on a conserved region of the 16S rRNA gene was used in conjunction with two different reverse primers specific for [A.] muris, targeting the 16S-23S internal transcribed spacer sequences. The specificity of the assay was tested against 78 reference and clinical isolates of Pasteurellaceae, including 37 strains of [A.] muris. In addition, eight other mice associated bacterial species which could pose a diagnostic problem were included. The assay showed 100% sensitivity and 97.95% specificity. Identification of the clinical isolates was validated by ITS profiling and when necessary by 16S rRNA sequencing. This multiplex PCR represents the first molecular tool able to detect [A.] muris and may become a reliable alternative to the present diagnostic methods.
authors.affiliation: Animal Research Facility, Heinrich-Heine-University Duesseldorf, University Hospital, Duesseldorf, Germany. Electronic address: Laurentiu.Benga@med.uni-duesseldorf.de.
authors.forename: Laurentiu
authors.initials: L
authors.lastname: Benga
authors.affiliation: Animal Research Facility, Heinrich-Heine-University Duesseldorf, University Hospital, Duesseldorf, Germany.
authors.forename: W Peter M
authors.initials: WPM
authors.lastname: Benten
authors.affiliation: Animal Research Facility, Heinrich-Heine-University Duesseldorf, University Hospital, Duesseldorf, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Engelhardt
authors.affiliation: Animal Research Facility, Heinrich-Heine-University Duesseldorf, University Hospital, Duesseldorf, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Gougoula
authors.affiliation: Animal Research Facility, Heinrich-Heine-University Duesseldorf, University Hospital, Duesseldorf, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Sager
mesh_terms: D000188:Actinobacillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D000189:Actinobacillus Infections / Q000382:microbiology / Q000662:veterinary*; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D012376:Rodent Diseases / Q000382:microbiology*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S
keywords: Internal transcribed spacer; Pasteurellaceae; Rodents; [Actinobacillus] muris