The ceja-1 sequence as a potential new molecular marker for Paracoccidioides brasiliensis infection.
We have developed a two-step PCR assay that amplifies a region of the ceja-1 sequence that is specific for virulent strains of Paracoccidioides brasiliensis. An internal region of the ceja-1 sequence was chosen for designing primers that were utilised in a single tube heminested PCR protocol to amplify DNA from six virulent strains. PCR specificity was determined by the absence of amplified products with genomic DNA from four non-virulent strains of P. brasiliensis and from eight fungal pathogens, one bacterium, two protozoa, one worm and mouse and human genomic DNA (leucocytes). The fact that the PCR product was only obtained with the genetic material from virulent isolates of P. brasiliensis suggested that this partial amplified sequence might be a marker of virulence for this fungus. The diagnostic potential of this PCR was confirmed by the successful amplification of this fragment with genomic DNA obtained in lymph node aspirate from a patient with paracoccidioidomycosis.
Correia
Janaina
J
Department of Immunology, Aggeu Magalhães Research Center, FIOCRUZ, Recife, PE, Brazil.
de Moraes Borba
Cíntia
C
Reis
Beatriz
B
Martins
Albert
A
Unkles
Sheila
S
Kinghorn
James Robert
JR
Lucena-Silva
Norma
N
D000818:Animals; D015415:Biomarkers; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009172:Mycology / Q000379:methods*; D010228:Paracoccidioides / Q000235:genetics* / Q000302:isolation & purification*; D010229:Paracoccidioidomycosis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D015415:Biomarkers; D017931:DNA Primers; D004271:DNA, Fungal