title: Detection of Mycoplasma mycoides subsp. mycoides SC in bronchoalveolar lavage fluids of cows based on a TaqMan real-time PCR discriminating wild type strains from an lppQ(-) mutant vaccine strain used for DIVA-strategies.
abstract: Contagious bovine pleuropneumonia (CBPP) is the most serious cattle disease in Africa, caused by Mycoplasma mycoides subsp. mycoides small-colony type (SC). CBPP control strategies currently rely on vaccination with a vaccine based on live attenuated strains of the organism. Recently, an lppQ(-) mutant of the existing vaccine strain T1/44 has been developed (Janis et al., 2008). This T1lppQ(-) mutant strain is devoid of lipoprotein LppQ, a potential virulence attribute of M. mycoides subsp. mycoides SC. It is designated as a potential live DIVA (Differentiating Infected from Vaccinated Animals) vaccine strain allowing both serological and etiological differentiation. The present paper reports on the validation of a control strategy for CBPP in cattle, whereby a TaqMan real-time PCR based on the lppQ gene has been developed for the direct detection of M. mycoides subsp. mycoides SC in ex vivo bronchoalveolar lavage fluids of cows and for the discrimination of wild type strains from the lppQ(-) mutant vaccine strain.
authors.affiliation: Research Unit, Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Länggass-Strasse 122, P.O. Box, 3001 Bern, Switzerland. edy.vilei@vbi.unibe.ch
authors.forename: Edy M
authors.initials: EM
authors.lastname: Vilei
authors.forename: Joachim
authors.initials: J
authors.lastname: Frey
mesh_terms: D000349:Africa; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D001428:Bacterial Vaccines; D001431:Bacteriological Techniques / Q000379:methods*; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D055786:Gene Knockout Techniques; D008969:Molecular Sequence Data; D009176:Mycoplasma mycoides / Q000302:isolation & purification*; D015345:Oligonucleotide Probes / Q000235:genetics; D011002:Pleuropneumonia, Contagious / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D037521:Virulence Factors / Q000172:deficiency
chemical_list: D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D037521:Virulence Factors
references.citation: Genome Res. 2004 Feb;14(2):221-7
references.pmid: 14762060
references.citation: Ann N Y Acad Sci. 2000;916:326-32
references.pmid: 11193642
references.citation: Vet J. 2007 Nov;174(3):513-21
references.pmid: 17157043
references.citation: Vet Microbiol. 2005 Nov 30;111(1-2):51-8
references.pmid: 16257143
references.citation: Microbiology (Reading). 2000 Feb;146 ( Pt 2):477-486
references.pmid: 10708386
references.citation: Vet Microbiol. 2004 Jun 3;100(3-4):283-8
references.pmid: 15145506
references.citation: Vet Microbiol. 2002 Jan 23;84(3):195-205
references.pmid: 11731172
references.citation: J Vet Diagn Invest. 1994 Oct;6(4):428-34
references.pmid: 7858022
references.citation: Clin Diagn Lab Immunol. 2001 Jan;8(1):85-92
references.pmid: 11139200
references.citation: Vet Res. 2003 Nov-Dec;34(6):761-75
references.pmid: 14746771
references.citation: Genomics. 2006 Nov;88(5):633-41
references.pmid: 16919417
references.citation: Mol Cell Probes. 1997 Apr;11(2):103-11
references.pmid: 9160324
references.citation: J Clin Microbiol. 1999 Dec;37(12):3815-21
references.pmid: 10565890
references.citation: Clin Diagn Lab Immunol. 2000 Jul;7(4):588-95
references.pmid: 10882657
references.citation: Dev Biol (Basel). 2003;114:147-60
references.pmid: 14677685
references.citation: N Z Vet J. 2008 Feb;56(1):40-7
references.pmid: 18322559
references.citation: BMC Microbiol. 2007 Apr 17;7:31
references.pmid: 17439646
references.citation: J Bacteriol. 1999 Feb;181(4):1319-23
references.pmid: 9973360
references.citation: Mol Cell Probes. 1999 Feb;13(1):23-8
references.pmid: 10024429
references.citation: Microbiology (Reading). 2008 Aug;154(Pt 8):2427-2436
references.pmid: 18667575
references.citation: Vet Microbiol. 1994 Dec;42(4):327-39
references.pmid: 9133058
references.citation: Rev Sci Tech. 2006 Dec;25(3):999-1011
references.pmid: 17361766
references.citation: Int J Syst Evol Microbiol. 2009 Jun;59(Pt 6):1353-8
references.pmid: 19502315
references.citation: Antonie Van Leeuwenhoek. 2008 Jan-Feb;93(1-2):175-83
references.pmid: 17674137
references.citation: BMC Microbiol. 2009 Oct 09;9:215
references.pmid: 19818124
references.citation: Vet Res. 2009 Jul-Aug;40(4):35
references.pmid: 19364458
references.citation: Vet Rec. 2002 Aug 3;151(5):156
references.pmid: 12199439
references.citation: Gene. 1995 Jul 4;160(1):95-100
references.pmid: 7628725
references.citation: Ann N Y Acad Sci. 2000;916:71-80
references.pmid: 11193704
references.citation: Infect Immun. 2008 Jan;76(1):263-9
references.pmid: 17998309
references.citation: Clin Microbiol Rev. 1994 Oct;7(4):533-58
references.pmid: 7834604
references.citation: J Bacteriol. 2005 Oct;187(19):6824-31
references.pmid: 16166545
references.citation: Comp Immunol Microbiol Infect Dis. 2010 Jul;33(4):279-90
references.pmid: 19187963
references.citation: J Vet Diagn Invest. 2006 Mar;18(2):168-71
references.pmid: 16617697
references.citation: J Mol Biol. 1990 Oct 5;215(3):403-10
references.pmid: 2231712
references.citation: Vet Microbiol. 2004 Mar 5;98(3-4):229-34
references.pmid: 15036531
references.citation: Mol Cell Probes. 2008 Oct-Dec;22(5-6):324-8
references.pmid: 18678244