title: Development and application of a real-time TaqMan(®) qPCR assay for detection and quantification of 'Candidatus Mycoplasma haemolamae' in South American camelids.
abstract: Two alpacas from a herd in southwest Switzerland died for unknown reasons. Necropsy revealed chronic weight loss and pale mucous membranes. Infection with hemotropic mycoplasmas was suspected and subsequently confirmed by molecular methods. In order to investigate the epidemiological situation in this herd, a real-time TaqMan(®) qPCR assay for the specific detection and quantification of hemoplasma infection in South American camelids was developed. This assay was based on the 16S rRNA gene and amplified 'Candidatus Mycoplasma haemolamae' DNA, but not DNA from other hemoplasmas or non-hemotropic mycoplasma species. The lower detection limit was one copy/PCR, and the amplification efficiency was 97.4%. In 11 out of 24 clinically healthy herd mates of the two infected alpacas, 'Candidatus M. haemolamae' infection was confirmed. No correlation was found between bacterial load and clinical signs or anemia. The assay described herein enables to detect and quantify 'Candidatus M. haemolamae' and may be used in future studies to investigate the prevalence, pathogenesis and treatment follow-up of hemoplasma infections in South American camelids.
authors.affiliation: Clinical Laboratory, Vetsuisse Faculty, University of Zurich, CH-8057 Zurich, Switzerland. mmeli@vetclinics.uzh.ch
authors.forename: Marina L
authors.initials: ML
authors.lastname: Meli
authors.forename: Christine
authors.initials: C
authors.lastname: Kaufmann
authors.forename: Patrik
authors.initials: P
authors.lastname: Zanolari
authors.forename: Nadia
authors.initials: N
authors.lastname: Robert
authors.forename: Barbara
authors.initials: B
authors.lastname: Willi
authors.forename: Hans
authors.initials: H
authors.lastname: Lutz
authors.forename: Regina
authors.initials: R
authors.lastname: Hofmann-Lehmann
mesh_terms: D000818:Animals; D002161:Camelids, New World / Q000382:microbiology*; D005260:Female; D008297:Male; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics*; D009175:Mycoplasma Infections / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013557:Switzerland
chemical_list: D012336:RNA, Ribosomal, 16S