RESUMO
Glutathione S-transferases (GSTs) are a group of phase II detoxification enzymes, which catalyze the conjugation of glutathione (GSH) with carcinogens, among other xenobiotics. The GSTM3 gene is part of the GSTs gene family, and its polymorphism A/B has been associated with risk and protective effects of several cancers. This genetic variant is a deletion of 3 bp (AGG) in intron 6. Previous association studies have performed genotyping using techniques such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In this study, we took advantage of the TaqMan(®) probes features and developed a reliable, faster, more simple and economic method to identify the 3-bp deletion. Our allelic discrimination method was able to distinguish between homozygous A/A, heterozygous A/B and homozygous B/B samples, as shown by TaqMan(®) based real-time PCR. Results were validated by Sanger Sequencing. In conclusion, we developed a specific and rapid method to detect the 3-bp deletion from the GSTM3 A/B polymorphism.
Assuntos
Glutationa Transferase/genética , Alelos , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Sondas de DNA , Genótipo , Heterozigoto , Homozigoto , Humanos , Íntrons , Isoenzimas/genética , Polimorfismo Genético , Reação em Cadeia da Polimerase em Tempo Real , Deleção de SequênciaRESUMO
Chromoblastomycosis is a subcutaneous, chronic, granulomatous mycosis that occurs more frequently in tropical and subtropical countries. We describe a case of a 63-year-old male patient with diagnosis of chromoblastomycosis by Cladophialophora carrionii with an 18-year evolution who developed a lethal squamous cell carcinoma.
Assuntos
Antifúngicos/uso terapêutico , Ascomicetos/genética , Cromoblastomicose/diagnóstico , Cromoblastomicose/tratamento farmacológico , DNA Espaçador Ribossômico/genética , Ascomicetos/efeitos dos fármacos , Ascomicetos/isolamento & purificação , Sequência de Bases , Carcinoma de Células Escamosas , Cromoblastomicose/microbiologia , DNA Fúngico/genética , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/tratamento farmacológico , Infecções Oportunistas/microbiologia , Análise de Sequência de DNA , Resultado do TratamentoRESUMO
We identified 29 Cladophialophora carrionii isolates recovered from Venezuelan patients with chromoblastomycosis using phenotypic and molecular characteristics. The genetic diversity of isolates was assessed by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) fingerprinting. We detected four electrophoretic patterns divided into two main clusters (I and II) comprising 10 and 17 isolates, respectively, and two minor clusters (III and IV) with one isolate each. An interesting cluster-age-lesion type association was detected. The median age of patients in cluster I was 37.5 years and in cluster II, 55 years of age (P = 0.04). The C. carrionii isolates found in cluster I were generally obtained from crusty lesions (60%) and isolates in cluster II were usually recovered from plaque type lesions (53%) even though the P values were only slightly less than significant (P = 0.08). No associations were found among the genetic features strains in the two clusters and gender, occupation, geographic origin, lesion size, severity, and duration of the disease. There was also no correlation between antifungal susceptibilities and strain clustering. In conclusion, molecular typing using ERIC-PCR revealed a genomic heterogeneity in the C. carrionii clinical isolates studied.
Assuntos
Antifúngicos/farmacologia , Ascomicetos/genética , Cromoblastomicose/microbiologia , Variação Genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Ascomicetos/efeitos dos fármacos , Ascomicetos/isolamento & purificação , Ascomicetos/fisiologia , Sequência de Bases , Criança , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Demografia , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Dados de Sequência Molecular , Sequências Repetitivas de Ácido Nucleico , Análise de Sequência de DNA , Venezuela , Adulto JovemRESUMO
A case of chromoblastomycosis of the hand caused by Rhinocladiella aquaspersa is described. The case was acquired locally in tropical Venezuela and was successfully treated with oral itraconazole.
