A novel real-time PCR assay for specific detection and quantification of Mycobacterium avium subsp. paratuberculosis in milk with the inherent possibility of differentiation between viable and dead cells

Dzieciol, Monika; Volgger, Patrick; Khol, Johannes; Baumgartner, Walter; Wagner, Martin; Hein, Ingeborg

doi:10.1186/1756-0500-3-251

Table 1 Bacterial strains used for inclusivity and exclusivity testing of the Mptb52.16 real-time PCR assay

From: A novel real-time PCR assay for specific detection and quantification of Mycobacterium avium subsp. paratuberculosis in milk with the inherent possibility of differentiation between viable and dead cells

Genus and species	Source
Mycobacterium avium subsp. paratuberculosis	CIP^a 103964
Mycobacterium avium subsp. paratuberculosis	CIP 103967
Mycobacterium avium subsp. paratuberculosis	CIP 103968
Mycobacterium avium subsp. paratuberculosis	CIP 103971
Mycobacterium avium subsp. paratuberculosis	CIP 103972
Mycobacterium avium subsp. paratuberculosis	CIP 103973
Mycobacterium avium subsp. paratuberculosis	CIP 103974
Mycobacterium avium subsp. paratuberculosis	CIP 103975
Mycobacterium avium subsp. paratuberculosis	CIP 103976
Mycobacterium avium subsp. paratuberculosis	CIP 103977
Mycobacterium abscessus	DSM^b 44196
Mycobacterium aurum	DSM 43999
Mycobacterium avium subsp.avium	DSM 44156
Mycobacterium avium subsp.silvaticum	DSM 44175
Mycobacterium chitae	DSM 44633
Mycobacterium duvalii	DSM 44244
Mycobacterium flavescens	DSM 43991
Mycobacterium fortuitum subsp. fortuitum	DSM 46621
Mycobacterium gordonae	DSM 44160
Mycobacterium intracellulare	DSM 43223
Mycobacterium kansasii	DSM 44162
Mycobacterium marinum	DSM 44344
Mycobacterium neoaurum	DSM 44074
Mycobacterium nonchromogenicum	DSM 44164
Mycobacterium parafortuitum	DSM 43528
Mycobacterium phlei	DSM 43239
Mycobacterium scrofulaceum	DSM 43992
Mycobacterium shiumoidei	DSM 44152
Mycobacterium thermoresistibile	DSM 44167
Mycobacterium triviale	DSM 44153
Mycobacterium vaccae	DSM 43292
Mycobacterium xenopi	DSM 43995
Bacillus cereus	NCTC^c 7464
Clostridium bovis	DSM 20582
Clostridium pyogenes	DSM 20630
Enterococcus faecalis	ATCC^d 19433
Escherichia coli	NCTC 9001
Lactococcus lactis	DSM 20069
Listeria monocytogenes	NCTC 11994
Pseudomonas aeruginosa	NCTC 10662
Salmonella typhimurium	ATCC 14028
Staphylococcus aureus	NCTC 1803
Staphylococcus epidermidis	ATCC 12228
Streptococcus agalactiae	DSM 2134
Streptococcus dysgalactiae subsp. dysgalactiae	DSM 20662
Streptococcus dysgalactiae subsp. equisimilis	DSM 6176
Streptococcus thermophilus	DSM 20617
Streptococcus uberis	DSM 20569

Colonies (MAP and non-MAP mycobacteria) suspended in 1 ml of Ringer's solution or overnight broth cultures (raw milk microflora) were subjected to DNA isolation using the NucleoSpin tissue kit (Machery-Nagel, Düren, Germany). The DNA concentration was measured fluorimetrically using a HoeferDyNA Quant200 apparatus (Pharmacia Biotech, San Francisco, USA) and the DNA was diluted to 0.1 ng/μl ddH₂O.
Notes: ^a CIP: Institute Pasteur, ^b DSM: German Collection of Microorgansims, ^c NCTC: National Collection of Type Cultures, ^d ATCC: American Type Culture Collection

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ISSN: 1756-0500

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