New probes used for IS1245 and IS1311 restriction fragment length polymorphism of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis isolates of human and animal origin in Norway

Abstract

BackgroundMycobacterium avium is an environmental mycobacterium that can be divided into the subspecies avium, hominissuis, paratuberculosis and silvaticum. Some M. avium subspecies are opportunistic pathogens for animals and humans. They are ubiquitous in nature and can be isolated from natural sources of water, soil, plants and bedding material. Isolates of M. avium originating from humans (n = 37), pigs (n = 51) and wild birds (n = 10) in Norway were examined by IS1245 and IS1311 RFLP using new and specific probes and for the presence of IS901 and ISMpa1 by PCR. Analysis and generation of a dendrogram were performed with the software BioNumerics.ResultsIS1311 RFLP provided clear results that were easy to interpret, while IS1245 RFLP generated more complex patterns with a higher discriminatory power. The combination of the two methods gave additional discrimination between isolates. All avian isolates except one were M. avium subsp. avium with two copies of IS1311 and one copy of IS1245, while the isolates of human and porcine origin belonged to M. avium subsp.hominissuis. The isolates from human patients were distributed randomly among the clusters of porcine isolates. There were few identical isolates. However, one isolate from a human patient was identical to a porcine isolate. Regional differences were detected among the porcine isolates, while there was no clustering of human isolates according to type of clinical symptoms or geographical location of the patient's home addresses.ConclusionThe results demonstrate that a wide range of M. avium subsp.hominissuis are present in pigs and humans in Norway, and that some of these isolates are very similar. It remains to be determined whether humans are infected from pigs or if they are infected from common environmental sources.

Non-Tuberculous Mycobacteria in Uganda: A Problem or Not?

“…This study showed that mycobacteriamia among febrile HIV-infected Ugandan adults accounted for 13% (Ssali et al, 1998). M. avium is also known to be one of the leading causes of infant lymphadenitis worldwide (Coetzer, 2004;Johansen, 2007;Van Ingen et al, 2009). This is in agreement with a study done in Uganda on the cause of cervical lymphadenitis.…”

Determination of Genotypic Diversity ofMycobacterium aviumSubspecies from Human and Animal Origins by Mycobacterial Interspersed Repetitive-Unit-Variable-Number Tandem-Repeat and IS1311Restriction Fragment Length Polymorphism Typing Methods

“…Whereas IS1311 RFLP usually generates clear hybridization patterns, IS1245 RFLP yields complex multiband patterns which are difficult to compare among different experiments and laboratories, mainly because of the heterogeneity in the intensities of the hybridization bands (19,20,42). Recently, an even simpler PCRbased molecular typing method, multilocus variable-number tandem-repeat analysis (MLVA), which is based on mycobacterial repetitive elements called mycobacterial interspersed repetitive-unit-variable-number tandem repeats (MIRU-VNTRs) (14,34,36,37), has been described for M. avium subsp.…”

“…For epidemiological investigations of MAC, the current reference molecular typing technique is restriction fragment length polymorphism analysis (RFLP) based on the IS1245 (47) and IS1311 (19,20) insertion sequences. Whereas IS1311 RFLP usually generates clear hybridization patterns, IS1245 RFLP yields complex multiband patterns which are difficult to compare among different experiments and laboratories, mainly because of the heterogeneity in the intensities of the hybridization bands (19,20,42).…”

Examination of Mycobacterium avium subsp. avium distribution in naturally infected hens by culture and triplex quantitative real time PCR

“…With the discovery of insertion elements in the mycobacterial genome, the determination of M. avium subspecies has since became more precise (Shamputa et al, 2004;Moravkova et al, 2008). MAA is characterised by the presence of 2 to 17 copies of the IS901 insertion sequence (Dvorska et al, 2003;Inglis et al, 2003) and a single copy of IS1245 (Johansen et al, 2007). To the authors' knowledge, no study for the specific identification of MAA in the tissue of naturally infected hens has previously been reported.…”