Development of a Species-Specific PCR-Restriction Fragment Length Polymorphism Analysis Procedure for Identification of
            <i>Madurella mycetomatis</i>

Ahmed, Abdallah E.; Mukhtar, Moawia M.; Kools-Sijmons, M.; Fahal, Ahmed Hassan; Hoog, Sybren de; Ende, Bert Gerrits van den; Zijlstra, E.E.; Verbrugh, Henri A.; Abugroun, El Sir A. M.; Elhassan, Ahmed M.; Belkum, Alex van

doi:10.1128/jcm.37.10.3175-3178.1999

Cited by 105 publications

(61 citation statements)

References 22 publications

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“…The infection is characterized by massive swelling and tumour formation, especially in the extremities. The source of the infectious Madurella mycetomatis strains, which are primarily clonal (Ahmed et al, 2003b), has long been enigmatic, and a search for the ecological niche was only possible after the development of molecular diagnostic tools for the species (Ahmed et al, 2003a). Despite the fact that the strains could not be cultured from soil and plant parts, these diagnostic tools very clearly indicated the presence of Madurella mycetomatis in the environment in soil samples and thorns from acacia trees (Ahmed et al, 2002).…”

Section: Pathogenicity Of Rhizopus Spp (Zygomycetes)mentioning

confidence: 99%

Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps?

et al. 2007

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It is common knowledge that pathogenic viruses can change hosts, with avian influenza, the HIV, and the causal agent of variant Creutzfeldt-Jacob encephalitis as well-known examples. Less well known, however, is that host jumps also occur with more complex pathogenic microorganisms such as bacteria and fungi. In extreme cases, these host jumps even cross kingdom of life barriers. A number of requirements need to be met to enable a microorganism to cross such kingdom barriers. Potential cross-kingdom pathogenic microorganisms must be able to come into close and frequent contact with potential hosts, and must be able to overcome or evade host defences. Reproduction on, in, or near the new host will ensure the transmission or release of successful genotypes. An unexpectedly high number of cross-kingdom host shifts of bacterial and fungal pathogens are described in the literature. Interestingly, the molecular mechanisms underlying these shifts show commonalities. The evolution of pathogenicity towards novel hosts may be based on traits that were originally developed to ensure survival in the microorganism's original habitat, including former hosts.

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Section: Pathogenicity Of Rhizopus Spp (Zygomycetes)mentioning

confidence: 99%

Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps?

et al. 2007

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“…Hence a rapid and accurate diagnostic tool to achieve the definitive species identification is considered a critical part in patient treatment and management [21–23]. Different laboratory techniques for species identification are in use, including culture, histophatology, [7, 25], FNAC [8, 24], serological assays and imaging [26 – 29] as well as different molecular diagnostic tools [30 – 35]. However, not all these assays are available in endemic regions.…”

Section: Discussionmentioning

confidence: 99%

The Accuracy of Histopathological and Cytopathological Techniques in the Identification of the Mycetoma Causative Agents

Fahal

Siddig

Mhmoud

et al. 2018

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24Mycetoma is a devastating neglected tropical disease, caused by various fungal 25 and bacterial pathogens. Correct diagnosis to the species level is mandatory for 26 proper treatment. In endemic areas, various diagnostic tests and techniques are 27 in use to achieve that, and that includes grain culture, surgical biopsy 28 histopathological examination, fine needle aspiration cytological (FNAC) 29 examination and in certain centres molecular diagnosis such as PCR. 30 In this retrospective study, the sensitivity, specificity and diagnostic accuracy of 31 grain culture, surgical biopsy histopathological examination and FNAC to identify 32 the mycetoma causative organisms were determined. The histopathological 33 examination appeared to have better sensitivity and specificity. 34The histological examination results were correct in 714 (97.5%) out of 750 35 patients infected with Madurella mycetomatis, in 133 (93.6%) out of 142 patients 36 infected with Streptomyces somaliensis, in 53 (74.6%) out of 71 patients infected 37 with Actinomadura madurae and in 12 (75%) out of 16 patients infected with 38 Actinomadura pelletierii. 39 FNAC results were correct in 604 (80.5%) out of 750 patients with Madurella 40 mycetomatis eumycetoma, in 50 (37.5%) out of 133 Streptomyces somaliensis 41 patients, 43 (60.5%) out of 71 Actinomadura madurae patients and 11 (68.7%) out 42 of 16 Actinomadura pelletierii. The mean time required to obtain the FNAC result 43was one day, and for the histopathological examinations results it was 3.5 days, 44 and for grain, it was a mean of 16 days. 45In conclusion, histopathological examination and FNAC are more practical 46 techniques for rapid species identification than grain culture in many endemic 47 regions. 48 49 50 Author summary 51 In mycetoma endemic regions, the medical and health settings are commonly 52 suboptimal, and only a few diagnostic tests and techniques are available. That had 53 badly affected the patients' proper diagnosis and management and thus the late 54presentation of patients with advanced disease. In this retrospective study, the 55 experience of the MRC on the common in use diagnostic tests in the period 56 between 1991 and 2018 is presented. 57In this study, the sensitivity, specificity rates and diagnostic accuracy of grain 58 culture, surgical biopsy histopathological examination and FNAC to identify the 59 mycetoma causative organisms were determined. The histopathological 60 examination appeared to have better sensitivity and specificity. Furthermore, the 61 grain culture identification needs high experience, it is the tedious procedure, and 62 cross-contamination is common hence misdiagnosis is frequent. It can be 63 concluded that histopathological examination and FNAC are more practical 64 techniques for rapid species identification than grain culture in many endemic 65 regions with poor diagnostic setting. 66 67 68 Introduction: 69 Mycetoma is a chronic granulomatous subcutaneous inflammatory infection 70 endemic in subtropical and tropical re...

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“…These fungal isolates were obtained from both the Mycetoma Research Center in Sudan and the Westerdijk Fungal Biodiversity Institute in the Netherlands and maintained in Erasmus Medical Centre. All isolates were identified to the species level on the basis of morphology, polymerase chain reaction (PCR)-based restriction fragment length polymorphisms, and sequencing of the ITS regions (3, 14, 21).…”

Section: Methodsmentioning

confidence: 99%

“…Madurella mycetomatis is a fungus only recognised as a pathogen in mycetoma and is responsible for more than 70% of all mycetoma infections in endemic areas (1, 12, 13). In 1999, specific PCR primers based on the internal transcribed spacer (ITS) region were developed for M. mycetomatis by our group (14), however, recently, this M. mycetomatis specific PCR primer pair was discovered to cross-react with Madurella pseudomycetomatis (15). Back then, M. pseudomycetomatis was not yet discovered (16).…”

Section: Introductionmentioning

confidence: 99%

The development of a novel diagnostic PCR forMadurella mycetomatisusing a comparative genome approach

Lim

Eadie

Siddig

et al. 2020

Preprint

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14 15 16 17 18 19 Corresponding author 20 Wendy WJ van de Sande 21 w.vandesande@erasmusmc.nl 22 23 Keyword: Mycetoma, Madurella mycetomatis, Comparative genome, Neglected tropical disease 24 25 26 ABSTRACT: 27 28 Eumycetoma is a neglected tropical disease characterized by large tumorous lesions. It is most 29 commonly caused by the fungus Madurella mycetomatis which accounts for more than 70% of cases 30 in central Africa. Currently, identification of the causative agent can only be reliably performed by a 31 species-specific PCR. However, we recently demonstrated that our M. mycetomatis specific PCR can 32 cross-react with Madurella pseudomycetomatis. We therefore used a comparative genome approach 33 to develop a new M. mycetomatis specific PCR for species identification. For this we compared the 34 published M. mycetomatis genome to genomes of other organisms in BLASTCLUST to identify unique 35 M. mycetomatis predicted protein coding sequences. Based on 16 of these unique sequences, PCR36 primers were developed. The specificity of these primers was further evaluated in other 37 eumycetoma causing agents including the Madurella sibling species. Out of the 16 tested sequences, 38 only one was unique for M. mycetomatis and this should be used as a novel diagnostic marker for M.39 mycetomatis. 40 41 42 43 INTRODUCTION: 44 45The neglected tropical disease mycetoma presents itself as a subcutaneous chronic granulomatous 46 infectious and inflammatory disease and is characterized by tumorous lesions (1, 2). This disease can 47 be caused by more than 70 different micro-organisms and is categorized into actinomycetoma 48 (caused by bacteria) and eumycetoma (caused by fungi). Most cases occur in the mycetoma belt 49 between the latitudes 15° South and 30° North. Diagnosis of eumycetoma is often only made 50 clinically in endemic areas due to the scarcity of facilities, expertise and financial capacity. 51Identification of the causative agent is time consuming and often limited to culture and histology 52 which can leads to misidentifications (1, 3, 4). The only way to properly identify eumycetoma 53 causative agents to the species level is through molecular identification, commonly PCR. PCR 54 identification's reliability, high turnover and sensitivity has made it widely used in diagnosing fungal 55 disease, and in most cases, it has already replaced culturing of the microorganism as the primary 56 diagnostic method (5-11). 58Madurella mycetomatis is a fungus only recognised as a pathogen in mycetoma and is responsible for 59 more than 70% of all mycetoma infections in endemic areas (1, 12, 13). In 1999, specific PCR primers 60 based on the internal transcribed spacer (ITS) region were developed for M. mycetomatis by our 61 group (14), however, recently, this M. mycetomatis specific PCR primer pair was discovered to cross-62 react with Madurella pseudomycetomatis (15). Back then, M. pseudomycetomatis was not yet 63 discovered (16). Since new fungi causing eumycetoma are still being discovered, there is clearly a 64 need...

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Development of a Species-Specific PCR-Restriction Fragment Length Polymorphism Analysis Procedure for Identification of Madurella mycetomatis

Cited by 105 publications

References 22 publications

Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps?

Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps?

The Accuracy of Histopathological and Cytopathological Techniques in the Identification of the Mycetoma Causative Agents

The development of a novel diagnostic PCR forMadurella mycetomatisusing a comparative genome approach

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