title: Real-time PCR assay for the detection of species of the genus Mannheimia.
abstract: Infections caused by species of the genus Mannheimia cause diverse disease complexes in many wild and domestic animals worldwide. Fast and accurate detection of single species within the genus remains an unsolved problem till today. To resolve this diagnostic challenge, we developed a real-time PCR assay for the rapid and specific identification of five species of the genus Mannheimia (M. haemolytica, M. varigena, M. ruminalis, M. granulomatis and M. glucosida) from bacterial cultures and tissue samples. The assay was validated with reference strains, field isolates and bacteria spiked tissue samples. The sodA gene was used as target region for species-specific primer pairs. The real-time PCR assay demonstrated species specificity for all five examined Mannheimia spp. and a rapid test completion time of less than 5 h. This is a considerable advantage compared to the traditional phenotyping methods currently used to distinguish between the species of the genus. The assay was able to detect approximately 10(3) bacterial cells per gram lung tissue sample, as determined with spiked tissue samples. We assume that the assay could become useful for fast laboratory diagnostic assessment particularly of respiratory infections caused by Mannheimia in animals.
authors.affiliation: Institut für Mikrobiologie und Tierseuchen, FU Berlin, Philippstr. 13, 10115 Berlin, Germany. guenther.sebastian@vetmed.fu-berlin.de
authors.forename: Sebastian
authors.initials: S
authors.lastname: Guenther
authors.forename: Peter
authors.initials: P
authors.lastname: Schierack
authors.forename: Mirjam
authors.initials: M
authors.lastname: Grobbel
authors.forename: Antina
authors.initials: A
authors.lastname: Lübke-Becker
authors.forename: Lothar H
authors.initials: LH
authors.lastname: Wieler
authors.forename: Christa
authors.initials: C
authors.lastname: Ewers
mesh_terms: D000820:Animal Diseases / Q000175:diagnosis / Q000382:microbiology*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002415:Cats; D002417:Cattle; D004285:Dogs; D031121:Mannheimia / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016871:Pasteurellaceae Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012756:Sheep; D013482:Superoxide Dismutase / Q000235:genetics; D013552:Swine
chemical_list: D001426:Bacterial Proteins; C095500:SodA protein, Bacteria; D013482:Superoxide Dismutase