A standardised restriction fragment length polymorphism (RFLP) method for typing Mycobacterium avium isolates links IS901 with virulence for birds

Dvorská, L.; Bull, Tim J.; Bartoš, Milan; Mátlová, L.; Svastova, Petra; Weston, Ross Tim; Kintr, Jaromir; Parmova, I.; Soolingen, Dick van; Pavlík, I.

doi:10.1016/s0167-7012(03)00092-7

Cited by 70 publications

(57 citation statements)

References 43 publications

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“…paratuberculosis (4,16), the cause of paratuberculosis (Johne's disease) in animals. IS901 is strongly associated with MAC infections in avian species, and IS901-positive strains are thus often called bird-type MAC members (7,8,39). IS1311 is consistently associated with the MAC, with only rare exceptions reported (2,21).…”

Section: Discussionmentioning

confidence: 99%

Efficient Differentiation ofMycobacterium aviumComplex Species and Subspecies by Use of Five-Target Multiplex PCR

et al. 2010

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Infections caused by the Mycobacterium avium complex (MAC) are on the rise in both human and veterinary medicine. A means of effectively discriminating among closely related yet pathogenetically diverse members of the MAC would enable better diagnosis and treatment as well as further our understanding of the epidemiology of these pathogens. In this study, a five-target multiplex PCR designed to discriminate MAC organisms isolated from liquid culture media was developed. This MAC multiplex was designed to amplify a 16S rRNA gene target common to all Mycobacterium species, a chromosomal target called DT1 that is unique to M. avium subsp. avium serotypes 2 and 3, to M. avium subsp. silvaticum, and to M. intracellulare, and three insertion sequences, IS900, IS901, and IS1311. The pattern of amplification results allowed determination of whether isolates were mycobacteria, whether they were members of the MAC, and whether they belonged to one of three major MAC subspecies, M. avium subsp. paratuberculosis, M. avium subsp. avium, and M. avium subsp. hominissuis. Analytical sensitivity was 10 fg of M. avium subsp. paratuberculosis genomic DNA, 5 to 10 fg of M. avium subsp. avium genomic DNA, and 2 to 5 fg of DNA from other mycobacterial species. Identification accuracy of the MAC multiplex was evaluated by testing 53 bacterial reference strains consisting of 28 different mycobacterial species and 12 nonmycobacterial species. Identification accuracy in a clinical setting was evaluated for 223 clinical MAC isolates independently identified by other methods. Isolate identification agreement between the MAC multiplex and these comparison assays was 100%. The novel MAC multiplex is a rapid, reliable, and simple assay for discrimination of MAC species and subspecies in liquid culture media.Since the early 1980s, there has been an increase in disease caused by organisms broadly categorized as nontuberculous mycobacteria (NTM), a generic term for mycobacteria not in the Mycobacterium tuberculosis complex and other than M. leprae (32). Of these NTM, Mycobacterium avium complex (MAC) species are the most common cause of human and animal disease globally (6,14,16,24). The clinical relevance of the MAC in humans has been amplified in recent decades with the increasing population of immunocompromised individuals resulting from longer life expectancy, immunosuppressive chemotherapy, and the AIDS pandemic (27 Members of the family Mycobacteriaceae, comprising the MAC, differ in virulence and ecology. Those designated M. avium subsp. hominissuis are genomically diverse, low-virulence, opportunistic pathogens for both animals and humans.

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Section: Discussionmentioning

confidence: 99%

Efficient Differentiation ofMycobacterium aviumComplex Species and Subspecies by Use of Five-Target Multiplex PCR

et al. 2010

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“…Among the most widely used methods has been the testing of species-specific insertion sequences. Despite the advantages of using these generally high-copy-number elements, there are at least 10 different insertion sequences found or characterized in MAC (8), some of them related, and cross-hybridization with PCR primers or probes used in restriction fragment length polymorphism (RFLP) procedures can potentially be misleading (14,16). In the case of M. avium subsp.…”

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Sequencing of hsp65 Distinguishes among Subsets of the Mycobacterium avium Complex

Turenne

Semret

Cousins³

et al. 2006

J Clin Microbiol

137

157

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The Mycobacterium avium complex consists of epidemiologically distinct subsets. The classification of these subsets is complicated by a number of factors, including the ambiguous results obtained with phenotypic and genetic assays and the recent appreciation that human and avian strains appear to be distinct. In previous work, sequencing based on a 441-bp portion of the hsp65 gene has proven to efficiently classify isolates within the Mycobacterium genus but provides low resolution for distinguishing among members of the M. avium complex. Therefore, in this study, we have targeted the more variable 3 region of the hsp65 gene to determine whether it can effectively discriminate M. avium complex isolates at the levels of species and subspecies. Primers designed for this target consistently generated amplicons for all organisms classified as M. avium complex. Sequences obtained indicate that M. intracellulare is genetically divergent from M. avium organisms, and distinct sequevars were obtained for M. avium subsets, including M. avium subsp. avium (bird type), M. avium subsp. hominissuis, and M. avium subsp. paratuberculosis. In addition, sequence differences served to distinguish bovine from ovine strains of M. avium subsp. paratuberculosis. A unique profile for M. avium subsp. silvaticum was not obtained. These results indicate that sequencing the 3 region of the hsp65 gene can simply and unambiguously distinguish species and subspecies of the M. avium complex.

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“…The most common causative agent found in tuberculous lesions from mesenteric, submandibular and occasionally inguinal ln of animals is M. a. hominissuis (MAH) of serotypes 4-6, 8-11 and 21 and of genotype dnaJ+, IS901-and IS1245+ (Leinemann et al, 1993;Morita et al, 1994;Nishimori et al, 1995;Balian et al, 1997;Ritacco et al, 1998;Dvorska et al, 1999Dvorska et al, , 2003Pavlik et al, 2000Pavlik et al, , 2003Ramasoota et al, 2001;Mijs et al, 2002). The sources of MAH, the causative agent of avian mycobacteriosis, are mostly various components of the life environment: water, soil, bedding, dust and invertebrates (Horvathova et al, 1997;Kazda, 2000;Pavlik et al, 2000;Fischer et al, 2001;Matlova et al, 2003;Trckova et al, 2004Trckova et al, , 2005Trckova et al, , 2006aSkoric et al, 2007).…”

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Isolation of conditionally pathogenic mycobacteria from the environment of one pig farm and the effectiveness of preventive measures between 1997 and 2003

Pavlík¹,

Mátlová²,

Gilar³

et al. 2007

Vet. Med. - Czech

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392Tuberculosis in pigs causes high economic losses farmers from all over the world. The most consequential causative agents of tuberculosis in pigs at present are members of Mycobacterium tuberculosis (MTC) and M. avium (MAC) complexes. The losses are above all caused by restriction of animal transport from infected farms (with the exception of slaughterhouses), culling of animals positive during the tuberculin testing and the price of meat and organs from slaughtered infected animals (Alfredsen and Skjerve 1993;Dey and Parham, 1993;Morita et al., 1994;Balian et. al., 1997;Cvetnic et al., 1998; Positive skin responses were found in animals with antibodies against MAH (18; 23.7%), MAA (3; 4.0%) and MI (9; 11.8%) antigens. By serological examination of 17 sows with repeated dubious responses for tuberculin skin testing with avian tuberculin, no antibodies against MAA were detected; MAH antibodies and MI antibodies were found in eight and two animals, respectively. By post mortem examination of lymph nodes (ln) and organ samples from all 76 animals with responses to avian tuberculin, no tuberculous/tuberculoid lesions were detected. By culture examination of ln and organs from 13 animals, conditionally pathogenic mycobacteria (CPM) were isolated from only one animal (breeding boar): from mesenteric, pulmonary, hepatic ln and from spleen tissue samples. These isolates were identified as MAH and CPM by the PCR method and biochemically. By investigation of the external environment (205 samples), 33 (16.3%) CPM isolates were obtained: 13 MAH, eight M. fortuitum, one M. nonchromogenicum, one M. abscessus, one M. scrofulaceum and nine unidentified isolates, which were non-MAC according to the PCR examination. Non-specific responses obtained in the intravital tests (skin and serological tests) caused by CPM present in the environment substantially complicated diagnosis of avian tuberculosis. Based on these findings, animal hygiene measures have been adopted since 2002; resulted in a decrease of environmental contamination with CPM and a reduction in the number of animals giving positive responses to avian tuberculin.

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A standardised restriction fragment length polymorphism (RFLP) method for typing Mycobacterium avium isolates links IS901 with virulence for birds

Cited by 70 publications

References 43 publications

Efficient Differentiation ofMycobacterium aviumComplex Species and Subspecies by Use of Five-Target Multiplex PCR

Efficient Differentiation ofMycobacterium aviumComplex Species and Subspecies by Use of Five-Target Multiplex PCR

Sequencing of hsp65 Distinguishes among Subsets of the Mycobacterium avium Complex

Isolation of conditionally pathogenic mycobacteria from the environment of one pig farm and the effectiveness of preventive measures between 1997 and 2003

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