RESUMEN
Trichophyton indotineae is an emerging species of the Trichophyton mentagrophytes complex (TMC), responsible for an epidemic of widespread hairless skin infections that is frequently (50-70%) resistant to terbinafine. In order to initiate appropriate treatment as quickly as possible without waiting for culture positivity (10-15 days) and molecular identification from the strain, we developed a dual quantitative PCR (qPCR) for the direct detection of T. indotineae in clinical samples. We first designed a T. indotineae-specific qPCR assay (TI-qPCR) targeting a single specific polymorphism in the internal transcribed spacer region. Although none of the 94 non-dermatophyte and 7 dermatophyte species were amplified, this TI-qPCR allowed amplification of other TMC species at a lower yield. With equal amounts (0.1 ng) of DNA per reaction, the mean quantitative cycle (Cq) values for T. indotineae and non-indotineae TMC were 27.9 (±0.1) and 38.9 (±0.3), respectively. Therefore, we normalized this assay against a previously validated pan-dermatophyte qPCR assay (PD-qPCR) and relied on the ΔCq [(TI-qPCR) - (PD-qPCR)] to identify T. indotineae versus other TMC species. Dual assay was validated using 86 clinical samples of culture-confirmed T. indotinea and 19 non-indotineae TMC cases. The mean ΔCq for non-indotineae TMC was 9.6 ± 2.7, whereas the ΔCq for T. indotinea was -1.46 ± 2.1 (P < .001). Setting the ΔCq at 4.5 as a cutoff value resulted in 100% specificity for the detection of T. indotineae. This dual qPCR assay quickly detects T. indotineae from skin scrapings, aiding in early diagnosis and treatment for patients with suspected infection.
Identifying the emerging species Trichophyton indotineae is long and requires to wait for culture positivity. We developed a dual qPCR strategy to detect T. indotineae directly from clinical sample with a 100% sensitivity.
Asunto(s)
Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y Especificidad , Tiña , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Tiña/diagnóstico , Tiña/microbiología , ADN de Hongos/genética , Trichophyton/genética , Trichophyton/aislamiento & purificación , Trichophyton/clasificación , Técnicas de Diagnóstico Molecular/métodos , ADN Espaciador Ribosómico/genéticaRESUMEN
Dermatophyte infections (a.k.a. ringworm, tinea) affect an estimated 20%-25% of the world's population. In North America, most dermatophytoses are caused by Trichophyton rubrum or Trichophyton mentagrophytes species complexes. Severe and antifungal-resistant dermatophytoses are a growing global public health problem. A new species of the T. mentagrophytes species complex, Trichophyton indotineae, has recently emerged and is notable for the severe infections it causes, its propensity for antifungal resistance, and its global spread. In this issue of the Journal of Clinical Microbiology, C. F. Cañete-Gibas, J. Mele, H. P. Patterson, et al. (J Clin Microbiol 61:e00562-23, 2023, https://doi.org/10.1128/JCM.00562-23) summarize the results of speciation and AFST performed on North American dermatophyte isolates received at a fungal diagnostic reference laboratory. Within their collection, 18.6% of isolates were resistant to terbinafine (a first-line oral antifungal for dermatophytoses), and similar proportions of T. rubrum and T. indotineae demonstrated terbinafine resistance. The authors also found that T. indotineae has been present in North America since at least 2017. These findings highlight the importance of increased surveillance efforts to monitor trends in severe and antifungal-resistant dermatophytoses and the need for antifungal stewardship efforts, the success of which is contingent upon improving laboratory capacity for dermatophyte speciation and AFST.
Asunto(s)
Arthrodermataceae , Tiña , Humanos , Terbinafina , Antifúngicos/farmacología , Arthrodermataceae/aislamiento & purificación , Tiña/tratamiento farmacológico , Tiña/epidemiología , Pruebas de Sensibilidad Microbiana , Trichophyton/clasificación , Farmacorresistencia Fúngica/efectos de los fármacosRESUMEN
BACKGROUND: During the past 5 years, an outbreak of recalcitrant dermatophytosis due to a novel Trichophyton species generally resistant to terbinafine, T. indotineae, has spread out from South Asia to many countries around the World. These isolates cannot be reliably differentiated from other Trichophyton spp. on the basis of morphological traits and the current laboratory diagnostics relies on sequencing of ribosomal DNA ITS region. OBJECTIVES: In this study, we aimed to introduce two inexpensive and rapid PCR-based assays for differentiation between T. indotineae and other dermatophytes. METHODS: The first introduced assay is based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, involving the amplification of TOP2 sequences and the digestion of PCR products by Cfr13I restriction enzyme. The second assay is proposed as conventional endpoint species-specific PCR amplification of the C120-287 intergenic locus. To validate the assays, a total of 191 Trichophyton spp. and 2 Microsporum canis strains with known ITS region sequences were used. From the T. mentagrophytes / T. interdigitale species complex (TMTISC), strains with 18 different ITS genotypes were tested. The sample of TMTISC isolates included 41 T. indotineae strains. RESULTS: TOP2 PCR-RFLP and T. indotineae-specific PCR were positive with testing on DNA of all 41 T. indotineae isolates and two strains of T. mentagrophytes belonging to ITS Types XIII and XVI, but negative with other species and other TMTISC ITS genotypes (n = 152). Therefore, the specificity of both new assays was 99%. CONCLUSION: The two developed diagnostic assays provide accurate and cost-effective means of identifying cultured T. indotineae isolates.
Asunto(s)
Tiña , Trichophyton , Humanos , ADN de Hongos/genética , ADN de Hongos/análisis , ADN Ribosómico , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Trichophyton/clasificación , Tiña/diagnóstico , Tiña/microbiologíaRESUMEN
Trichophyton interdigitale, an anthropophilic species, is one of the main causative agents of tinea unguium and tinea pedis. T. interdigitale and the zoophilic species T. mentagrophytes are morphologically and physiologically very similar. Isolates of the T. interdigitale/T. mentagrophytes complex from around the world have been classified into more than 10 internal transcribed spacer (ITS) genotypes. In this study, we isolated T. interdigitale from Japanese patients and investigated which ITS type was more common. The ITS regions of 29 clinical isolates of T. interdigitale and one clinical isolate of T. mentagrophytes were sequenced. The phylogenetic analysis of the ITS region sequences revealed that the 29 isolates of T. interdigitale belong to ITS type II of T. interdigitale. The one clinical isolate of T. mentagrophytes was in the same cluster with ITS type II* of T. mentagrophytes. One terbinafine-resistant strain of T. interdigitale also belonged to ITS type II of T. interdigitale.
Asunto(s)
Trichophyton , Humanos , Pueblos del Este de Asia , Filogenia , Trichophyton/clasificación , Trichophyton/aislamiento & purificación , ADN Espaciador Ribosómico/genéticaRESUMEN
Arthroderma is the most diverse genus among dermatophytes encompassing species occurring in soil, caves, animal burrows, clinical material and other environments. In this study, we collected ex-type, reference and authentic strains of all currently accepted Arthroderma species and generated sequences of three highly variable loci (ITS rDNA, ß-tubulin, and translation elongation factor 1-α). The number of accepted species was expanded to 27. One novel species, A. melbournense (ex-type strain CCF 6162T = CBS 145858T), is described. This species was isolated from toenail dust collected by a podiatrist in Melbourne, during an epidemiological study of four geographical regions of Eastern Australia. Trichophyton terrestre, Chrysosporium magnisporum, and Chrysosporium oceanitis are transferred to Arthroderma. Typification is provided for T. terrestre that is not conspecific with any of the supposed biological species from the former T. terrestre complex, that is, A. insingulare, A. lenticulare and A. quadrifidum. A multi-gene phylogeny and reference sequences provided in this study should serve as a basis for future phylogenetic studies and facilitate species identification in practice. LAY ABSTRACT: The genus Arthroderma encompasses geophilic dermatophyte species that infrequently cause human and animal superficial infections. Reference sequences from three genetic loci were generated for all currently accepted Arthroderma species and phylogeny was constructed. Several taxonomic novelties are introduced. The newly provided data will facilitate species identification and future taxonomic studies.
Asunto(s)
Arthrodermataceae/clasificación , Arthrodermataceae/genética , ADN de Hongos/genética , Genes Fúngicos/genética , Filogenia , Australia , ADN Espaciador Ribosómico/genética , Humanos , Microsporum/clasificación , Microsporum/genética , Factor 1 de Elongación Peptídica/genética , Trichophyton/clasificación , Trichophyton/genética , Tubulina (Proteína)/genéticaRESUMEN
BACKGROUND: Tinea capitis is a common dermatophyte infection of the scalp primarily affecting children, with less frequent, though not rare, observation in adults. OBJECTIVE: This study aimed to assess changes in the causative agents of adult tinea capitis over a 60-year period in the Hubei area. METHODS: A retrospective, single-centre study was performed on 164 adults with tinea capitis between 1960 and 2020. RESULTS: Out of 1113 cases of tinea capitis, 164 patients were adults, representing 14.7% of all patients. Adult tinea capitis was slightly more prevalent in males (91, 55.5%) than in females (73, 44.5%), but gender difference was not statistically significant between adults and children. Adult tinea capitis was most prevalent between the ages of 18 and 29 years, with a mean age of 22 years. Trichophyton schoenleinii was the most common dermatophyte in adult tinea capitis (78, 47.6%), followed by Trichophyton violaceum (58, 35.4%). Most adult tinea capitis cases before the 1980s were caused by T. schoenleinii, but T. violaceum has become the leading pathogen for recent adult tinea capitis cases. CONCLUSION: Tinea capitis is not a disease exclusive to children. On the contrary, an upward trend of tinea capitis in adults has been observed in recent years. T. violaceum has become the dominant causative agent of adult tinea capitis in the Hubei area, replacing T. schoenleinii. These results provide a better understanding of the treatment and prevention of tinea capitis in adults.
Asunto(s)
Tiña del Cuero Cabelludo , Trichophyton/clasificación , Adolescente , Adulto , China/epidemiología , Femenino , Humanos , Masculino , Estudios Retrospectivos , Cuero Cabelludo , Tiña del Cuero Cabelludo/epidemiología , Tiña del Cuero Cabelludo/microbiología , Adulto JovenRESUMEN
Background: The Trichophyton mentagrophytes complex is the second most common causal agent of dermatophytosis. It comprises five species-T. mentagrophytes, T. interdigitale, T. erinacei, T quinckeanum, and T. benhamie, as well as nine different genotypes of T. mentagrophytes / T. interdigitale-which are morphologically similar; however, their susceptibility to antifungal agents may differ. For targeted therapy and better prognosis, it is important to identify these species at a molecular level. However, since many hospitals lack molecular methods, the actual aetiology of dermatophytosis caused by this complex remains unknown. Objective: To characterize 55 anthropophilic isolates of the T. mentagrophytes complex recovered from a dermatological centre in Yucatán, Mexico. Material and methods: Fifty-five isolates of the T. mentagrophytes complex were obtained from patients with tinea capitis, tinea pedis, tinea corporis, tinea barbae, and tinea unguium. They were characterized by their colonial and microscopic morphology on Sabouraud dextrose agar (SDA) and through the sequencing of a fragment from the region ITS1-5.8S-ITS2. Results: All colonies grown on SDA were white. Forty-six isolates formed colonies with a powdery texture, while nine isolates formed colonies with a velvety texture. The micromorphological features were typical of the T. mentagrophytes complex. The molecular analysis revealed that 55 isolates were microorganisms that belonged to the T. mentagrophytes complex, that 46 formed powdery colonies representing T. mentagrophytes, and that the other nine isolates that formed velvety colonies represented T. interdigitale. The latter nine isolates were obtained from patients with tinea pedis, tinea corporis, and tinea unguium. Conclusions: The colony morphology on SDA led to the identification of 46 isolates as T. mentagrophytes and nine isolates as T. interdigitale. At a molecular level, the species identified by their morphology were identified only as T. mentagrophytes complex.
Asunto(s)
Antifúngicos/farmacología , ADN Intergénico/genética , Tiña/genética , Trichophyton/genética , Dermatosis Facial/genética , Dermatosis Facial/microbiología , Genotipo , Humanos , Onicomicosis/genética , Onicomicosis/microbiología , Análisis de Secuencia de ADN , Tiña/microbiología , Tiña/patología , Tiña del Cuero Cabelludo/genética , Tiña del Cuero Cabelludo/microbiología , Tiña del Pie/genética , Tiña del Pie/microbiología , Trichophyton/clasificación , Trichophyton/efectos de los fármacos , Trichophyton/patogenicidadRESUMEN
BACKGROUND: Dermatophytoses rank among the most frequent communicable diseases in humans, and the zoonotic transmission is increasing. The zoophilic dermatophyte Trichophyton (T.) benhamiae is nowadays one of the main causes of tinea faciei et corporis in children. However, scientific data on molecular pathomechanisms and specific virulence factors enabling this ubiquitous occurrence are scarce. OBJECTIVES: To study tissue invasion and the expression of important virulence factors of T. benhamiae, isolates that were recovered from two groups of hosts (humans vs. guinea pigs (GP)) using an ex vivo skin model. METHODS: After confirmation of species identity by ITS sequencing, CFU suspensions of dermatophyte isolates (n = 20) were applied to the skin infection model and cultured. Employing specific immunofluorescence staining techniques, the expression of subtilisin 3 and 6 and metallocarboxypeptidase A was analysed. The general mode of invasion was explored. Results were compared with biopsies of naturally infected GP. RESULTS: All isolates were successfully recovered and proliferated well after application to the infection model. Progressive invasion of hyphae through all skin structures and destruction of explants were observed with early events being comparable to natural infection. An increasing expression of the examined virulence factors towards the end of culture was noticed but no difference between the two groups of isolates. CONCLUSIONS: For the first time, important in vivo markers of dermatophytosis were visualised immunohistochemically in an ex vivo skin infection model and in skin biopsies of GP naturally infected with T. benhamiae. More research on the underlying pathomechanisms of dermatophyte infection is urgently needed.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Piel/microbiología , Tiña/microbiología , Trichophyton/crecimiento & desarrollo , Trichophyton/patogenicidad , Factores de Virulencia , Animales , Modelos Animales de Enfermedad , Cobayas , Humanos , Técnicas de Cultivo de Órganos/métodos , Piel/patología , Trichophyton/clasificación , Percepción VisualRESUMEN
BACKGROUND: Dermatophytosis is one of the most frequent superficial mycoses in the world. MAIN AIM: To describe the cases of skin dermatophytosis and its main aetiologic agents in patients referred to a Mycological Reference Laboratory in Medellín, Colombia. METHODS: A retrospective study was carried out with records of patients referred between 1994 and 2016 to the Corporación para Investigaciones Biológicas (CIB), Medellín-Colombia, because of clinical suspicion of skin dermatophytosis. RESULTS: Of a total of 5628 clinical records of patients with suspicion of skin dermatophytosis analysed, 2780 (49.4%) had a proven or probable dermatophytosis diagnosis, 2774 cultures were performed, and aetiologic agents were isolated in 2576 samples (92.9%). The most frequently isolated aetiologic agents were Trichophyton rubrum (44.3%), followed by Trichophyton mentagrophytes complex (33.3%), Epidermophyton floccosum (12.4%), Nannizzia gypseum complex (5.7%, formerly Microsporum gypseum), Microsporum canis (3.5%) and Trichophyton tonsurans (0.8%). The most frequent clinical forms were tinea pedis (72.7%) and tinea corporis (12.7%). In addition, a group of patients (0.9%) developed mixed infections by two dermatophyte agents and another (4.1%) developed infections in more than one anatomical site. CONCLUSIONS: The results of the present study are coherent with previous reports where T rubrum and T mentagrophytes complex were the main causative agents of dermatophytosis. However, the increased incidence of N gypsea complex over M canis is worth highlighting.
Asunto(s)
Dermatomicosis/epidemiología , Hongos/aislamiento & purificación , Tiña/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Arthrodermataceae/clasificación , Arthrodermataceae/aislamiento & purificación , Niño , Preescolar , Colombia/epidemiología , Dermatomicosis/microbiología , Epidermophyton/clasificación , Epidermophyton/aislamiento & purificación , Hongos/clasificación , Humanos , Lactante , Masculino , Microsporum/clasificación , Microsporum/aislamiento & purificación , Persona de Mediana Edad , Estudios Retrospectivos , Piel/microbiología , Piel/patología , Tiña/epidemiología , Trichophyton/clasificación , Trichophyton/aislamiento & purificación , Adulto JovenRESUMEN
The anthropophilic dermatophyte Trichophyton tonsurans and its zoophilic counterpart T. equinum are phylogenetically closely related. The barcoding marker rDNA internal transcribed spacer (ITS) shows limited variation between these two species. In the current study, we combined molecular approaches with phenotypic data to determine the species boundaries between T. tonsurans (n = 52) and T. equinum (n = 15) strains originating from humans (n = 40), horses (n = 26), and a mouse (n = 1). Culture characteristics and physiology on Trichophyton agar media 1 and 5 were evaluated. Multi-locus sequencing involving ITS, partial large rDNA subunit (LSU), ß-tubulin (TUB), 60S ribosomal protein (RPB), and translation elongation factor-3 (TEF3) genes, and the mating-type (MAT) locus was performed. Amplified fragment length polymorphism data were added. None of the test results showed complete mutual correspondence. With the exception of strains from New Zealand, strains of equine origin required niacin for growth, whereas most strains from human origin did not show this dependence. It is concluded that T. tonsurans and T. equinum incompletely diverged from a common lineage relatively recently. MAT1-1 and MAT1-2 are the main distinguishing genes between the two species.
Asunto(s)
ADN Ribosómico/genética , Trichophyton/genética , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Animales , Biodiversidad , Genes del Tipo Sexual de los Hongos/genética , Genes del Tipo Sexual de los Hongos/fisiología , Caballos , Humanos , Ratones , Tipificación de Secuencias Multilocus , Trichophyton/clasificaciónRESUMEN
The most important species of the Trichophyton rubrum group are T. rubrum, causing mainly skin and nail infections, and T. violaceum which is mostly scalp-associated. The status of a third species, T. soudanense, has been under debate. With a polyphasic approach, using molecular phylogenetic techniques, MALDI-TOF mass spectrometry and physiological and morphological analysis, we re-evaluated the T. rubrum complex. Our results support four genetic lineages within the complex each with a distinct morphology and identifiable via MALDI-TOF MS: T. rubrum, T. violaceum, T. soudanense and the T. yaoundei clade. However, ITS and Bt2 sequencing data could not confirm these taxa as four monophyletic species. Our results also suggest that strains formerly identified as T. kuryangei and T. megninii should be considered in future taxonomic studies.
Asunto(s)
Trichophyton/clasificación , Trichophyton/genética , Arthrodermataceae/clasificación , Arthrodermataceae/genética , Filogenia , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Phylogenetic studies of the family Arthrodermataceae have revealed seven monophyletic dermatophyte clades representing the genera Trichophyton, Epidermophyton, Nannizzia, Lophophyton, Paraphyton, Microsporum, and Arthroderma. Members of the genus Nannizzia are geo- or zoophiles that occasionally infect humans. With the newly proposed taxonomy, the genus Nannizzia comprises thirteen species, i.e., Nannizzia aenigmatica, N. corniculata, N. duboisii, N. fulva, N. graeserae, N. gypsea, N. nana, N. incurvata, N. perplicata, N. persicolor, N. praecox, and two novel species. Nannizzia polymorpha sp. nov. was isolated from a skin lesion of a patient from French Guiana. For the strain originally described as Microsporum racemosum by Borelli in 1965, we proposed Nannizzia lorica nom. nov. The species are fully characterized with five sequenced loci (ITS, LSU, TUB2, RP 60S L1 and TEF3), combined with morphology of the asexual form and physiological features. A key to the species based on phenotypic and physiological characters is provided.
Asunto(s)
Arthrodermataceae/genética , Arthrodermataceae/clasificación , Epidermophyton/clasificación , Epidermophyton/genética , Microsporum/clasificación , Microsporum/genética , Filogenia , Trichophyton/clasificación , Trichophyton/genéticaRESUMEN
The Trichophyton rubrum species complex comprises commonly encountered dermatophytic fungi with a worldwide distribution. The members of the complex usually have distinct phenotypes in culture and cause different clinical symptoms, despite high genome similarity. In order to better delimit the species within the complex, molecular, phenotypic, and physiological characteristics were combined to reestablish a natural species concept. Three groups, T. rubrum, T. soudanense, and T. violaceum, could be distinguished based on the sequence of the internal transcribed spacer (ITS) ribosomal DNA barcode gene. On average, strains within each group were similar by colony appearance, microscopy, and physiology, but strains between groups showed significant differences. Trichophyton rubrum strains had higher keratinase activity, whereas T. violaceum strains tended to be more lipophilic; however, none of the phenotypic features were diagnostic. The results of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and amplified fragment length polymorphism (AFLP) were partially consistent with the ITS data but failed to distinguish the species unambiguously. Despite their close similarity, T. violaceum, T. soudanense, and T. rubrum can be regarded as independent species with distinct geographical distributions and clinical predilections. Trichophyton soudanense is pheno- and genotypically intermediate between T. rubrum and T. violaceum For routine diagnostics, ITS sequencing is recommended.
Asunto(s)
Trichophyton/clasificación , Análisis del Polimorfismo de Longitud de Fragmentos Amplificados , Análisis por Conglomerados , ADN de Hongos/química , ADN de Hongos/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Humanos , Técnicas Microbiológicas , Microscopía , Filogenia , Análisis de Secuencia de ADN , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tiña/microbiología , Trichophyton/genética , Trichophyton/aislamiento & purificación , Trichophyton/fisiologíaAsunto(s)
Antifúngicos , ADN de Hongos , Farmacorresistencia Fúngica , Análisis de Secuencia de ADN , Tiña , Trichophyton , Humanos , Tiña/microbiología , Tiña/tratamiento farmacológico , Tiña/diagnóstico , Trichophyton/genética , Trichophyton/efectos de los fármacos , Trichophyton/aislamiento & purificación , Trichophyton/clasificación , Antifúngicos/farmacología , Antifúngicos/uso terapéutico , Farmacorresistencia Fúngica/genética , ADN de Hongos/genética , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana EdadRESUMEN
There has been a considerable upsurge of extensive, treatment recalcitrant, dermatophytosis presenting as tinea corporis and tinea cruris in India since the past few years. Genome analysis of Trichophytonspecies causing severe superficial dermatophytosis in North India confirmed a unique clade related to the T.mentagrophytes/interdigitale complex, seeming to belong to an early diverging clade of the complex. The Indian Trichophyton species genomes were highly related showing only up to 42 SNPs between any two isolates confirming their clonal origin. Other genetic approaches such as ITS sequencing and multigene phylogeny used in this study were contradictory or inconclusive to show the differentiation of these isolates from T. mentagrophytes/T. interdigitale. Remarkably, high rates of resistance to all three commonly used oral antifungals, i.e., 36% for terbinafine (MICs 4 to ≥32â¯mg/L), 39.5% for fluconazole (MIC range 32 to ≥64â¯mg/L) and griseofulvin (Geometric mean MICâ¯≥â¯4â¯mg/L) were observed. Two important amino acid substitutions (Leu393Phe or Phe397Leu) leading to a terbinafine resistant phenotype were found in the squalene epoxidase protein of all tested terbinafine resistant isolates. All 20 examined genomes presented a high mobility group (HMG) domain transcription factor gene corresponding to mating type (+). Of these, three isolates also showed positivity for both alpha-box and HMG in the genome which might indicate hybridization or an incomplete sexual cycle. Therefore, we highlight the potential of this organism to rapidly spread alleles that might be driving antifungal resistance among its population. This new population of Trichophyton with high rates of in vitro antifungal resistance seems to be driving an ongoing outbreak of dermatophytosis in India. Our study highlights difficulties in identifying isolates from the Trichophyton mentagrophytes/interdigitale clade of the genus using currently available molecular tools. High resistance rates of terbinafine warrant further clinical studies to assess its utility in the treatment of dermatophytosis caused by this clonal strain.
Asunto(s)
Antifúngicos/farmacología , Farmacorresistencia Fúngica , Tiña/microbiología , Trichophyton/efectos de los fármacos , Brotes de Enfermedades , Resistencia a Múltiples Medicamentos , Genoma Fúngico , Humanos , India/epidemiología , Pruebas de Sensibilidad Microbiana , Tipificación Molecular , Tiña/epidemiología , Trichophyton/clasificación , Trichophyton/genéticaRESUMEN
The fungi Trichophyton mentagrophytes and T. interdigitale are closely related species, causing superficial infections in humans and other mammals. The status of these taxa is a field of long-lasting debates. To clarify their phylogenetic relationships within the genus Trichophyton and sharpen the species boundaries, we performed sequencing of four T. mentagrophytes genomes and also evaluated three previously published multilocus data sets. We performed computational species delimitation analysis on all available in GenBank internal transcribed spacer region (ITS) sequences of Trichophyton spp. Phylogenomic data, phylogenetic network, and species delimitation analyses implied that T. mentagrophytes and T. interdigitale belong to the same phylogenetic species. However, we argue that taxonomic status quo should be retained, from the perspective of epidemiological data and the principle of taxonomic stability. Since there is a correlation between ITS genotype and epidemiological source of an isolate, restriction of T. interdigitale to purely anthropophilic ITS genotypes seems to be reasonable.
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Genotipo , Filogenia , Trichophyton/clasificación , Biología Computacional , ADN de Hongos/genética , ADN Espaciador Ribosómico/genética , Genoma Fúngico , Genómica , Análisis de Secuencia de ADN , Secuenciación Completa del GenomaRESUMEN
Mating experiments were conducted with four clinical Trichophyton benhamiae isolates, genetically similar to the Trichophyton benhamiae CBS 112371, featuring the plus mating type and with two minus type strains. One minus type strain belonged to the white subgroup, and the other minus type strain, DSM 6916, showed genetic kinship to the yellow subgroup. Only two plus type strains were able to form mature, pigmented gymnothecia with DSM 6916. These two plus type strains demonstrated dark pigmentation and powdery mycelium on Takashio agar, whereas the other three strains exhibited a low degree of pigmentation on the same medium. All five plus strains were able to mate with the minus type strain of their own white subgroup. Cultures from single ascospore isolates showed highly variable morphology and pigmentation. Three genetic markers (ITS, mating type, EF1 alpha) were analyzed in polymerase chain reaction (PCR) experiments with optimized primers and PCR conditions to discriminate between subgroups. Furthermore, RAPD-PCR was used to generate a DSM 6916-specific DNA-fragment which served as an additional genetic marker. Assessing the isolates with recombinant genotypes, it was found that three genetic markers behave like linked genes. The recombination of plus mating type went together with ITS, EF1 alpha and RAPD marker of the DSM 6916 parental strain and was most frequently isolated, whereas plus types recombinants in this case were completely missing. This shows a high imbalance in mating type distribution of recombinants.
Asunto(s)
Genes del Tipo Sexual de los Hongos , Trichophyton/clasificación , Trichophyton/genética , ADN de Hongos/genética , ADN Intergénico/genética , Proteínas Fúngicas/genética , Marcadores Genéticos , GenotipoRESUMEN
The fungi Trichophyton mentagrophytes and T interdigitale account for significant amount of dermatophytosis cases worldwide. These two dermatophytes form a species complex and have a number of ribosomal internal transcribed spacer (ITS) region genotypes, allowing simultaneous species identification and strain typing. Our aim was to describe the geographic distribution of T mentagrophytes/T interdigitale ITS region genotypes and find an association between the genotypes and clinical presentations of respective infections. We performed rDNA ITS region sequencing in 397 Iranian T mentagrophytes/T interdigitale isolates and analysed all available in GenBank entries with sequences of this kind. For the study, 515 clinical annotations were available. Statistical analysis was performed by chi-squared test and Spearman rank correlation analysis. A total of 971 sequences belonged to genotypes with at least 10 geographic annotations and were classified on the basis of exclusive occurrence in a particular region or high relative contribution to a regional sample. We discerned Asian and Oceanian ("KU496915" Type V, "KT192500" Type VIII, "KU315316"), European ("FM986750" Type III, "MF926358" Type III*, "KT285210" Type VI) and cosmopolitan ("FM986691" Type I, "JX122216" Type II, "KP132819" Type II* and "AF170453" Type XXIV) genotypes. There was statistically significant difference in the ITS genotype distribution between different affected body sites. Trichophyton mentagrophytes "KT192500" Type VIII correlated with tinea cruris, T mentagrophytes "KU496915" Type V correlated with tinea corporis, T interdigitale "JX122216" Type II correlated with tinea pedis and onychomycosis. Trichophyton mentagrophytes and T interdigitale genotypes can be associated with distinct geographic locations and particular clinical presentations.
Asunto(s)
Genotipo , Filogenia , Tiña/microbiología , Trichophyton/clasificación , Trichophyton/genética , África , Américas , Asia , ADN de Hongos/genética , ADN Ribosómico , ADN Espaciador Ribosómico/genética , Europa (Continente) , Geografía , Humanos , Análisis de Secuencia de ADNRESUMEN
The disease burden of chronic-relapsing and therapy-refractory superficial dermatophytosis dramatically increased in India within the past 5-6 years. In order to evaluate the prevalence of this trend, 201 skin scrapings were collected from patients from all parts of India and were tested for dermatophytes using both fungal culture and a PCR-ELISA directly performed with native skin scrapings. Fungal culture material was identified by genomic Sanger sequencing of the internal transcribed spacer (ITS) region and the translation elongation factor (TEF)-1α gene. In total, 149 (74.13%) out of the 201 samples showed a dermatophyte-positive culture result. Out of this, 138 (92.62%) samples were identified as Trichophyton (T.) mentagrophytes and 11 (7.38%) as Trichophyton rubrum. The PCR-ELISA revealed similar results: 162 out of 201 (80.56%) samples were dermatophyte-positive showing 151 (93.21%) T mentagrophytes- and 11 (6.79%) T rubrum-positive samples. In this study, we show for the first time a dramatic Indian-wide switch from T rubrum to T mentagrophytes. Additionally, sequencing revealed a solely occurring T mentagrophytes "Indian ITS genotype" that might be disseminated Indian-wide due to the widespread abuse of topical clobetasol and other steroid molecules mixed with antifungal and antibacterial agents.
Asunto(s)
Epidemias , Tiña/epidemiología , Trichophyton/clasificación , Trichophyton/aislamiento & purificación , Adolescente , Adulto , Anciano , Niño , Preescolar , ADN de Hongos/química , ADN de Hongos/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , India/epidemiología , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Factor 1 de Elongación Peptídica/genética , Reacción en Cadena de la Polimerasa , Prevalencia , Análisis de Secuencia de ADN , Trichophyton/genética , Adulto JovenRESUMEN
An alarming pan Indian increase in the incidence of superficial dermatophytosis has been noticed over the past 5-6 years. Recent studies have demonstrated emerging predominance of Trichophyton (T.) mentagrophytes as the causative organism in such cases. Interestingly, a distinct Indian genotype of T. mentagrophytes has been identified and recognised with the help of sequencing of the ITS region of the rDNA. That has, however, led to a basic confusion owing to the newly introduced taxonomy of dermatophytes in 2017. According to this most recently suggested classification and new taxonomy of dermatophytes, the former "T. mentagrophytes complex" is differentiated into T. mentagrophytes (zoophilic strains) and T. interdigitale (anthropophilic strains). We have noticed that in some recent studies the causative agent of the chronic, relapsing dermatophytosis outbreak in India has been described as T. interdigitale. In our opinion, it is very likely that these T. interdigitale strains isolated in Delhi and Chennai in India are indeed strains more closely related to the neotype of T. mentagrophytes and not strains of T. interdigitale. We therefore want to underscore the importance of a common nomenclature of species in accordance with the new taxonomy of dermatophytes. This would most likely facilitate better understanding of the issue amongst dermatologists and microbiologists in general. Mistaken identification of Trichophyton isolates not limited to India is very likely to occur due to the lack of appropriate molecular diagnosis which in turn is based on the already published data that presumably wrongly identify one species instead of the other.