Mycobacterium avium complex (MAC) currently comprises eight species of environmental and animal-associated, slowly-growing mycobacteria: Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium colombiense, Mycobacterium arosiense, Mycobacterium bouchedurhonense, Mycobacterium marseillense and Mycobacterium timonense. In humans, MAC organisms are responsible for opportunistic infections whose unique epidemiology remains poorly understood, in part due to the lack of a genotyping method applicable to all eight MAC species. In this study we developed multispacer sequence typing (MST), a sequencing-based method, for the genotyping of MAC organisms. An alignment of the genome sequence of M. avium subsp. hominissuis strain104 and M. avium subsp. paratuberculosis strain K-10 revealed621 intergenic spacers <1000 bp. From these, 16 spacers were selected that ranged from 300 to 800 bp and contained a number of variable bases, <50 within each of the 16 spacers. Four spacers were successfully PCR-amplified and sequenced in 11 reference strains. Combining the sequence of these four spacers in 106 MAC organisms, including 83 M. avium, 11 M. intracellulare, six M. chimaera, two M. colombiense and one each of M. arosiense, M. bouchedurhonense, M. marseillense and M. timonense, yielded a total of 45 spacer types, with an index of discrimination of 0.94. Each spacer type was specific for a species and certain spacer types were specific for subspecies of M. avium. MST is a new method for genotyping of organisms belonging to any one of the eight MAC species tested in this study.
Ben SalahI.,
CayrouC.,
RaoultD.,
DrancourtM.2009; Mycobacterium marseillense sp. nov., Mycobacterium timonense sp.nov. and Mycobacterium bouchedurhonense sp. nov.,novel species in the Mycobacterium avium complex. Int J Syst Evol Microbiol 59:2803–2808
BietF.,
BoschiroliM. L.,
ThorelM. F.,
GuilloteauL. A.2005; Zoonotic aspects of Mycobacterium bovis and Mycobacterium avium-intracellulare complex (MAC. Vet Res 36:411–436
BillsN. D.,
HinrichsS. H.,
AdenT. A.,
WickertR. S.,
IwenP. C.2009; Molecular identification of Mycobacterium chimaera as a cause of infection in a patient with chronic obstructive pulmonary disease. Diagn Microbiol Infect Dis 63:292–295
Bruijnesteijn van CoppenraetL. E.,
de HaasP. E.,
LindeboomJ. A.,
KuijperE. J.,
van SoolingenD.2008; Lymphadenitis in children is caused by Mycobacterium avium hominissuis and not related to ‘bird tuberculosis’. Eur J Clin Microbiol Infect Dis 27:293–299
CilliersF. J.,
WarrenR. M.,
HaumanJ. H.,
WiidI. J.,
van HeldenP. D.1997; Oligonucleotide (GTG)5 as an epidemiological tool in the study of nontuberculous mycobacteria. J Clin Microbiol 35:1545–1549
CovertT. C.,
RodgersM. R.,
ReyesA. L.,
StelmaG. N.Jr1999; Occurrence of nontuberculous mycobacteria in environmental samples. Appl Environ Microbiol 65:2492–2496
Du MoulinG. C.,
StottmeierK. D.,
PelletierP. A.,
TsangA. Y.,
Hedley-WhyteJ.1988; Concentration of Mycobacterium avium by hospital hot water systems. JAMA 260:1599–1601
FalkinhamJ. O.III,
NortonC. D.,
LeChevallierM. W.2001; Factors influencing numbers of Mycobacterium avium, Mycobacterium intracellulare, and other mycobacteria in drinking water distribution systems. Appl Environ Microbiol 67:1225–1231
GuerreroC.,
BernasconiC.,
BurkiD.,
BodmerT.,
TelentiA.1995; A novel insertion element from Mycobacterium avium, IS 1245, is a specific target for analysis of strain relatedness. J Clin Microbiol 33:304–307
HilbornE. D.,
YakrusM. A.,
CovertT. C.,
HarrisS. I.,
DonnellyS. F.,
SchmittM. T.,
ToneyS.,
BaileyS. A.,
StelmaG. N.Jr2008; Molecular comparison of Mycobacterium avium isolates from clinical and environmental sources. Appl Environ Microbiol 74:4966–4968
HunterP. R.,
GastonM. A.1988; Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol 26:2465–2466
KasaiH.,
EzakiT.,
HarayamaS.2000; Differentiation of phylogenetically related slowly growing mycobacteria by their gyrB sequences. J Clin Microbiol 38:301–308
KomijnR. E.,
de HaasP. E.,
SchneiderM. M.,
EgerT.,
NieuwenhuijsJ. H.,
van den HoekR. J.,
BakkerD.,
van Zijd ErveldF. G.,
van SoolingenD.1999; Prevalence of Mycobacterium avium in slaughter pigs in The Netherlands and comparison of IS 1245 restriction fragment length polymorphism patterns of porcine and human isolates. J Clin Microbiol 37:1254–1259
KyriakopoulosA. M.,
TassiosP. T.,
Matsiota-BernardP.,
MarinisE.,
TsaousidouS.,
LegakisN. J.1997; Characterization to species level of Mycobacterium avium complex strains from human immunodeficiency virus-positive and -negative patients. J Clin Microbiol 35:3001–3003
Le DantecC.,
DuguetJ. P.,
MontielA.,
DumoutierN.,
DubrouS.,
VincentV.2002; Occurrence of mycobacteria in water treatment lines and in water distribution systems. Appl Environ Microbiol 68:5318–5325
MazurekG. H.,
HartmanS.,
ZhangY.,
BrownB. A.,
HectorJ. S.,
MurphyD.,
WallaceR. J.Jr1993; Large DNA restriction fragment polymorphism in the Mycobacterium avium- M. intracellulare complex: a potential epidemiologic tool. J Clin Microbiol 31:390–394
MijsW.,
de HaasP.,
RossauR.,
van der LaanL.,
RigoutsL.,
PortaelsF.,
van SoolingenD.2002; Molecular evidence to support a proposal to reserve the designation Mycobacterium avium subsp. avium for bird-type isolates and ‘ M.avium subsp. hominissuis’ for the human/porcine type of M. avium. Int J Syst Evol Microbiol 52: 1505. 1518
MoritaY.,
MaruyamaS.,
KabeyaH.,
NagaiA.,
KozawaK.,
KatoM.,
NakajimaT.,
MikamiT.,
KatsubeY.,
KimuraH.2004; Genetic diversity of the dnaJ gene in the Mycobacterium avium complex. J Med Microbiol 53:813–817
MurciaM. I.,
TortoliE.,
MenendezM. C.,
PalenqueE.,
GarciaM. J.2006; Mycobacterium colombiense sp. nov., a novel member of the Mycobacterium avium complex and description of MAC-X as a new ITS genetic variant. Int J Syst Evol Microbiol 56:2049–2054
NishiuchiY.,
MaekuraR.,
KitadaS.,
TamaruA.,
TaguriT.,
KiraY.,
HiragaT.,
HirotaniA.,
YoshimuraK.other authors2007; The recovery of Mycobacterium avium-intracellulare complex (MAC) from the residential bathrooms of patients with pulmonary MAC. Clin Infect Dis 45:347–351
Pestel-CaronM.,
GraffG.,
BerthelotG.,
PonsJ. L.,
LemelandJ. F.1999; Molecular analysis of Mycobacterium avium isolates by using pulsed-field gel electrophoresis and PCR. J Clin Microbiol 37:2450–2455
SantosR.,
OliveiraF.,
FernandesJ.,
GonçalvesS.,
MacieiraF.,
CadeteM.2005; Detection and identification of mycobacteria in the Lisbon water distribution system. Water Sci Technol 52:177–180
ThompsonJ. D.,
GibsonT. J.,
PlewniakF.,
JeanmouginF.,
HigginsD. G.1997; The CLUSTAL_X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882
TortoliE.,
RindiL.,
GarciaM. J.,
ChiaradonnaP.,
DeiR.,
GarzelliC.,
KroppenstedtR. M.,
LariN.,
MatteiR.other authors2004; Proposal to elevate the genetic variant MAC-A, included in the Mycobacterium avium complex, to species rank as Mycobacterium chimaera sp. nov. Int J Syst Evol Microbiol 54:1277–1285
TurenneC. Y.,
SemretM.,
CousinsD. V.,
CollinsD. M.,
BehrM. A.2006; Sequencing of hsp65 distinguishes among subsets of the Mycobacterium avium complex. J Clin Microbiol 44:433–440
TurenneC. Y.,
CollinsM.,
AlexanderD. C.,
BehrM. A.2008; Mycobacterium avium subsp. paratuberculosis and M. avium subsp. avium are independently evolved pathogenic clones of a much broader group of M. avium organisms. J Bacteriol 190:2479–2487