Comparison of DNA Microarray, Loop-Mediated Isothermal Amplification (LAMP) and Real-Time PCR with DNA Sequencing for Identification of Fusarium spp. Obtained from Patients with Hematologic Malignancies

Autor(es):Mostrar menos -

de Souza, Marcela ; Matsuzawa, Tetsuhiro ; Sakai, Kanae ; Muraosa, Yasunori ; Lyra, Luzia ; Busso-Lopes, Ariane Fidelis ; Shafferman Levin, Anna Sara ; Schreiber, Angelica Zaninelli ; Mikami, Yuzuru ; Gonoi, Tohoru ; Kamei, Katsuhiko ; Moretti, Maria Luiza ; Trabasso, Plinio

• Tipo de documento: Artigo Científico
• Fonte: Mycopathologia; v. 182, n. 7-8, p. 625-632, AUG 2017.
• Citações Web of Science: 2

Resumo

The performance of three molecular biology techniques, i.e., DNA microarray, loop-mediated isothermal amplification (LAMP), and real-time PCR were compared with DNA sequencing for properly identification of 20 isolates of Fusarium spp. obtained from blood stream as etiologic agent of invasive infections in patients with hematologic malignancies. DNA microarray, LAMP and real-time PCR identified 16 (80%) out of 20 samples as Fusarium solani species complex (FSSC) and four (20%) as Fusarium spp. The agreement among the techniques was 100%. LAMP exhibited 100% specificity, while DNA microarray, LAMP and real-time PCR showed 100% sensitivity. The three techniques had 100% agreement with DNA sequencing. Sixteen isolates were identified as FSSC by sequencing, being five Fusarium keratoplasticum, nine Fusarium petroliphilum and two Fusarium solani. On the other hand, sequencing identified four isolates as Fusarium non-solani species complex (FNSSC), being three isolates as Fusarium napiforme and one isolate as Fusarium oxysporum. Finally, LAMP proved to be faster and more accessible than DNA microarray and real-time PCR, since it does not require a thermocycler. Therefore, LAMP signalizes as emerging and promising methodology to be used in routine identification of Fusarium spp. among cases of invasive fungal infections. (AU)

• Processo FAPESP: 11/16205-5 - Identificação das espécies de Fusarium de amostras clínicas do Hospital de Clínicas da UNICAMP através das técnicas de Microarranjo de DNA e amplificação isotérmica em ciclos (LAMP).
• Beneficiário: Marcela de Souza
• Modalidade de apoio: Bolsas no Brasil - Mestrado