rDNA-RFLP identification of Candida species in immunocompromised and seriously diseased patients.
PCR was used to amplify a targeted region of the ribosomal DNA of 76 Candida spp. isolates from immunocompromised and seriously diseased patients. Thirty-seven strains isolated from different anatomical sites of 11 patients infected with HIV (Vitória, ES, Brazil), 26 isolates from patients under treatment at Odilon Behrens Hospital and 13 isolates from skin and urine samples from São Marcos Clinical Analysis Laboratory (Belo Horizonte, Brazil) were scored. Fragments of rDNA were amplified using primer pairs ITS1-ITS4, for the amplification of ITS1 and ITS2 regions, including the gene for the 5.8 s subunit. Amplification resulted in fragments ranging in size from 350 to 950 bp. Amplicons were digested with eight restriction enzymes. A pattern of species-specificity among the different medically important Candida species could be identified following restriction digestion of the PCR products. Candida albicans was the species most frequently observed, except for the group of newborns under treatment at the Odilon Behrens Hospital and for the isolates from the clinical analysis laboratory. C. parapsilosis was the species most frequently observed in these two groups.
Pinto
Patrícia M
PM
Departament de Microbiologia--Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Resende
Maria A
MA
Koga-Ito
Cristiane Y
CY
Ferreira
José A G
JA
Tendler
Miriam
M
D002175:Candida / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002176:Candida albicans / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D002177:Candidiasis / Q000382:microbiology*; D016000:Cluster Analysis; D016172:DNA Fingerprinting; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D021903:DNA, Ribosomal Spacer / Q000302:isolation & purification / Q000378:metabolism; D020459:Genes, rRNA; D006801:Humans; D016867:Immunocompromised Host; D017720:Molecular Epidemiology; D016533:Mycological Typing Techniques; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D012340:RNA, Ribosomal, 5.8S; D004262:DNA Restriction Enzymes