New restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus

Semighini, Camile P.; Delmas, Guillaume; Park, Steven; Amstrong, Donald; Perlin, David S.; Goldman, Gustavo H.

doi:10.1016/s0928-8244(01)00232-2

Cited by 8 publications

(11 citation statements)

References 11 publications

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“…Secondly, all PCR products derived from the various M. mycetomatis isolates presented the same restriction pattern after digestion by six different enzymes, which is in line with at least species homogeneity. That PCR-RFLP can be useful to detect fungal genetic variability has been shown repeatedly as well (5,14,17,24). To test whether the different primer combinations were species specific, they were tested on different fungal species.…”

Section: Discussionmentioning

confidence: 77%

Madurella mycetomatisStrains from Mycetoma Lesions in Sudanese Patients Are Clonal

et al. 2003

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Molecular diversity among clinical isolates of Madurella mycetomatis, the prime fungal agent of human mycetoma in Sudan, could possibly explain the diverse clinical presentations of this severely debilitating infectious disease. In addition, culture-independent DNA-mediated typing tests need to be developed for this organism, since M. mycetomatis DNA, but not the organism itself, can be identified in soil, the material from which infections are thought to originate. A collection of 38 different clinical M. mycetomatis isolates was characterized by large-scale random amplification of polymorphic DNA using 20 different primer species. These analyses, involving at least 2,600 annealing sites, showed a complete lack of DNA fingerprint variation among the various isolates. From the resulting homogeneous DNA fingerprints, seven fragments were cloned and sequenced, and novel, species-specific PCR restriction fragment length polymorphism (RFLP) tests were designed. The seven PCR RFLP tests were successfully performed on the 38 different M. mycetomatis strains. However, again all M. mycetomatis DNA patterns obtained appeared to be identical, whereas patterns produced using DNAs from other fungal species were clearly discriminatory. These results suggest that there is little genetic variation among clinically relevant M. mycetomatis strains from Sudan. The data tentatively imply that different manifestations of mycetoma are due to differences in host susceptibility rather than differential virulence of the causative agent.Mycetoma presents as a chronic, relatively painless, subcutaneous granulomatous lesion which is characterized by the formation of multiple sinuses. In ϳ40% of all infections, a fungus is the causative agent, the sclerotia of which are shed through these sinuses in the form of fungal grains (10, 13). The color of these grains has diagnostic value. Green grains identify Aspergillus flavus as the most likely causative agent, whereas white grains are usually produced by Pseudallescheria boydii, Aspergillus nidulans, or Acremonium kiliense. Brownish grains are produced by Neotestudina rosatii, while deep-black grains are produced by species such as Curvularia lunata, Exophiala jeanselmei, Pyrenochaeta romeroi, Leptosphaeria senegalensis, Madurella grisea, and Madurella mycetomatis. The last fungus is the most prevalent mycetoma agent in Sudan (10). Although this agent has been shown to occur in various geographic regions, most cases of M. mycetomatis mycetoma occur in a relatively comprehensive "mycetoma belt" (1). The precise mechanism of infection remains enigmatic, but it is frequently suggested that traumatic inoculation of fungus-containing soil, assisted by the presence of plant materials such as thorns, provides a likely route of inoculation (10,11,13,19). However, successful cultivation of the organism from soil has been documented only sparsely, although recent molecular detection has revealed that fungal DNA can be detected quite easily in soils from different regions in the mycetoma belt (1). Becau...

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Section: Discussionmentioning

confidence: 77%

Madurella mycetomatisStrains from Mycetoma Lesions in Sudanese Patients Are Clonal

et al. 2003

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“…To achieve finer discrimination of A. fumigatus strains, some authors have used different methods in combination (1,42,45). However, applying multiple tests makes the task of typing A. fumigatus more laborious, and some typing methods may be compromised in their accuracy, for example, because interpretation of band sizes is open to error.…”

Section: Discussionmentioning

confidence: 99%

“…One study even drew the conclusion of a neg-VOL. 45,2007 A. FUMIGATUS MLST 1475 on May 12, 2018 by guest http://jcm.asm.org/ ative association between clinical and environmental isolates (43). By contrast, two studies found indistinguishable isolates in several patients, a situation compatible with cross-infection with a common strain type (12,27); the finding is also more likely to arise in studies based on typing methods with low discriminatory power.…”

Section: Discussionmentioning

confidence: 99%

Multilocus Sequence Typing of the Pathogenic Fungus Aspergillus fumigatus

et al. 2007

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A multilocus sequence typing (MLST) scheme was devised for Aspergillus fumigatus. The system involved sequencing seven gene fragments and was applied to a panel of 100 isolates of A. fumigatus from diverse sources. Thirty different sequence types were found among the 100 isolates, and 93% of the isolates differed from the other isolates by only one allele sequence, forming a single clonal cluster as indicated by the eBURST algorithm. The discriminatory power of the MLST method was only 0.93. These results strongly indicate that A. fumigatus is a species of a relatively recent origin, with low levels of sequence dissimilarity. Typing methods based on variable numbers of tandem repeats offer higher levels of strain discrimination. Mating type data for the 100 isolates showed that 71 isolates were type MAT1-2 and 29 isolates were MAT1-1.

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“…Other defective retrotransposons, including Afut2 [45], Af4 , Af4A [46], and Taf1 [47] could also be used successfully for strain typing in A. fumigatus. In the recent paper on sequencing of the A. fumigatus genome, several retrotransposons have been identified, about half of them belonging to the gypsy family of retrotransposons with long terminal repeats [48].…”

Section: Rflp Analysismentioning

confidence: 99%

Molecular typing of aspergilli: Recent developments and outcomes

Varga

2006

Med Mycol

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Aspergillus spp. have been the subject of numerous epidemiological studies. The most useful typing techniques are DNA based methods including the random amplified polymorphic DNA technique, microsatellite length polymorphisms, restriction fragment length polymorphism (RFLP) analysis using retrotransposon-like sequences as probes, and multilocus sequence typing. The results of typing clinical isolates indicate that most of the invasive aspergillosis (IA) patients were infected by a single strain. Genetic analysis could not discriminate between clinical and environmental isolates of Aspergillus fumigatus, indicating that every strain present in the environment is a potential pathogen if it encounters the appropriate host. The source of infection can also be monitored by typing. Typing studies led to the discovery of a new pathogenic species, A. lentulus, and to the identification of several species not known previously to be pathogenic. Typing studies revealed the existence of two genetically isolated groups within a global A. fumigatus population. Aspergillus fumigatus was found to be the first example of a true cosmopolitan fungus. Additionally, the results obtained in several studies support the premise that recombination played an important role in A. fumigatus populations. The discovery of functional mating type genes in A. fumigatus indicates that past or recent sexual processes could be responsible for the observed recombining population structure.

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New restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus

Cited by 8 publications

References 11 publications

Madurella mycetomatisStrains from Mycetoma Lesions in Sudanese Patients Are Clonal

Madurella mycetomatisStrains from Mycetoma Lesions in Sudanese Patients Are Clonal

Multilocus Sequence Typing of the Pathogenic Fungus Aspergillus fumigatus

Molecular typing of aspergilli: Recent developments and outcomes

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