Rapid and selective detection of macrocyclic trichothecene producing Stachybotrys chartarum strains by loop-mediated isothermal amplification (LAMP)

Köck, Johannes; Gottschalk, Christoph; Ulrich, Sebastian; Schwaiger, Karin; Gareis, M.; Niessen, Ludwig

doi:10.1007/s00216-021-03436-y

Cited by 8 publications

(6 citation statements)

References 56 publications

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“…The A-type strain IBT 40288 possesses a complete AC1 cluster, and our previously published PCR data suggest the same for all H-type strains analyzed so far [11]. Further data strongly indicate that H-type strains are genetically heterogeneous and often lack one or more of the SC1 and/or SC3 genes [11,47]. This loss of genetic information can be construed as an erosive process, in which H-type strains derive from a strain harboring intact SC1 and SC3 and developed towards strains that lack these genetic elements.…”

Section: Discussionsupporting

confidence: 54%

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The Evolution of the Satratoxin and Atranone Gene Clusters of Stachybotrys chartarum

Ulrich

Lang

Niessen

et al. 2022

JoF

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Stachybotrys chartarum is frequently isolated from damp building materials or improperly stored animal forage. Human and animal exposure to the secondary metabolites of this mold is linked to severe health effects. The mutually exclusive production of either satratoxins or atranones defines the chemotypes A and S. Based upon the genes (satratoxin cluster, SC1-3, sat or atranone cluster, AC1, atr) that are supposed to be essential for satratoxin and atranone production, S. chartarum can furthermore be divided into three genotypes: the S-type possessing all sat- but no atr-genes, the A-type lacking the sat- but harboring all atr-genes, and the H-type having only certain sat- and all atr-genes. We analyzed the above-mentioned gene clusters and their flanking regions to shed light on the evolutionary relationship. Furthermore, we performed a deep re-sequencing and LC-MS/MS (Liquid chromatography–mass spectrometry) analysis. We propose a first model for the evolution of the S. chartarum genotypes. We assume that genotype H represents the most ancient form. A loss of the AC1 and the concomitant acquisition of the SC2 led to the emergence of the genotype S. According to our model, the genotype H also developed towards genotype A, a process that was accompanied by a loss of SC1 and SC3.

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Section: Discussionsupporting

confidence: 54%

“…This is true for the A-and S-type but also for H-type strains [11]. In a study comprising 105 S. chartarum strains, none possessed or lacked both the AC1 and the SC2 [47]. This strongly suggests that AC1 and SC2 are mutually exclusive.…”

Section: Discussionmentioning

confidence: 97%

The Evolution of the Satratoxin and Atranone Gene Clusters of Stachybotrys chartarum

Ulrich

Lang

Niessen

et al. 2022

JoF

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“…cultures came from the authors reference collection (see further details in Table S1).The 53 cultures of P. destructans that were used in the current study were isolated from bat swabs, bat guano or bat hibernacula environments (sediments/walls) from various locations in Europe and North America [ 8 , 15 , 20 , 35 ]. The self-isolated fungal strains shown in table S1 (marked in grey) were identified to species/clades level using micromorphology, FTIR spectroscopy or genetic methods (LAMP assay [ 36 , 37 ], gene sequencing, e.g. 28S rRNA, ITS, beta tubulin, calmodulin, or multi locus sequencing [ 15 ]).…”

Section: Methodsmentioning

confidence: 99%

Development and Application of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Diagnosis of the Bat White-Nose Disease Fungus Pseudogymnoascus destructans

et al. 2022

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Pseudogymnoascus destructans (= Geomyces destructans) is a psychrophilic filamentous fungus that causes White-Nose Disease (WND; the disease associated with White-Nose Syndrome, WNS) in hibernating bats. The disease has caused considerable reductions in bat populations in the USA and Canada since 2006. Identification and detection of the pathogen in pure cultures and environmental samples is routinely based on qPCR or PCR after DNA isolation and purification. Rapid and specific direct detection of the fungus in the field would strongly improve prompt surveillance, and support control measures. Based on the genes coding for ATP citrate lyase1 (acl1) and the 28S-18S ribosomal RNA intergenic spacer (IGS) in P. destructans, two independent LAMP assays were developed for the rapid and sensitive diagnosis of the fungus. Both assays could discriminate P. destructans from 159 tested species of filamentous fungi and yeasts. Sensitivity of the assays was 2.1 picogram per reaction (pg/rxn) and 21 femtogram per reaction (fg/rxn) for the acl1 and IGS based assays, respectively. Moreover, both assays also work with spores and mycelia of P. destructans that are directly added to the master mix without prior DNA extraction. For field-diagnostics, we developed and tested a field-applicable version of the IGS-based LAMP assay. Lastly, we also developed a protocol for preparation of fungal spores and mycelia from swabs and tape liftings of contaminated surfaces or infected bats. This protocol in combination with the highly sensitive IGS-based LAMP-assay enabled sensitive detection of P. destructans from various sources.

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“…Due to the fact that color and ornamentation of conidia change with age [15,37], it is easy to misidentify S. chartarum for such species as S. chlorohalonata, S. yunnanensis, Stachybotrys albipes and Stachybotrys elegans. For this reason, molecular biology techniques are an alternative solution enabling unambiguous identification of species of Stachybotrys taxa [38][39][40][41][42][43], especially those recently established based on fast and reliable MALDI-TOF MS identification method [44,45]. Sometimes, however, even well-designed specific primers used in real-time PCR procedure do not differentiate S. chartarum from S. chlorohalonata or S. yunnanensis [15].…”

Section: Taxonomic Positionmentioning

confidence: 99%

Update on Stachybotrys chartarum—Black Mold Perceived as Toxigenic and Potentially Pathogenic to Humans

Dyląg

Spychała

Zielinski

et al. 2022

Biology

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In nature, there are many species of fungi known to produce various mycotoxins, allergens and volatile organic compounds (VOCs), as well as the commonly known etiological agents of various types of mycoses. So far, none of them have provoked so much emotion among homeowners, builders, conservators, mycologists and clinicians as Stachybotrys chartarum. This species compared to fungi of the genera Fusarium and Aspergillus is not as frequently described to be a micromycete that is toxigenic and hazardous to human and animal health, but interest in it has been growing consistently for three decades. Depending on the authors of any given review article, attention is focused either on the clinical aspects alongside the role of this fungus in deterioration of biomaterials, or aspects related to its biology, ecology and taxonomic position. On the one hand, it is well established that inhalation of conidia, containing the highest concentrations of toxic metabolites, may cause serious damage to the mammalian lung, particularly with repeated exposure. On the other hand, we can find articles in which authors demonstrate that S. chartarum conidia can germinate and form hyphae in lungs but are not able to establish an effective infection. Finally, we can find case reports that suggest that S. chartarum infection is linked with acute pulmonary hemorrhage, based on fungal structures recovered from patient lung tissue. New scientific reports have verified the current state of knowledge and note that clinical significance of this fungus is exceedingly controversial. For these reasons, understanding S. chartarum requires reviewing the well-known toxigenic features and harmful factors associated with this fungus, by gathering the newest ones into a coherent whole. The research problem related to this fungus seems to be not overly publicized, and there is still a demand to truthfully define the real threats of S. chartarum and phylogenetically related species. The most important problem, which should be fully elucidated as soon as possible, remains the clarification of the pathogenicity of S. chartarum and related species. Maybe it is urgent time to ask a critical question, namely what exactly do we know 28 years after the outbreak of pulmonary hemorrhage in infants in Cleveland, Ohio, USA most likely caused by S. chartarum?

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Rapid and selective detection of macrocyclic trichothecene producing Stachybotrys chartarum strains by loop-mediated isothermal amplification (LAMP)

Cited by 8 publications

References 56 publications

The Evolution of the Satratoxin and Atranone Gene Clusters of Stachybotrys chartarum

The Evolution of the Satratoxin and Atranone Gene Clusters of Stachybotrys chartarum

Development and Application of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Diagnosis of the Bat White-Nose Disease Fungus Pseudogymnoascus destructans

Update on Stachybotrys chartarum—Black Mold Perceived as Toxigenic and Potentially Pathogenic to Humans

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