Detection of Candida albicans in blood by PCR in a rabbit animal model of disseminated candidiasis.
A model of acute disseminated Candida albicans infection in New Zealand rabbits was developed to determine the sensitivity and accuracy of polymerase chain reaction (PCR) assay compared with the lysis-centrifugation blood culture method. Primers used amplify a DNA fragment from the multicopy gene coding for the small subunit rRNA, highly conserved in fungi. The sensitivity of PCR achieved in rabbit blood samples spiked with Candida albicans was 10-50 CFU/100 microL. A nested-PCR increased the limit of detection 10-fold. The sensitivity achieved exclusively with the lysis-centrifugation method (37.5%) was higher than that obtained with PCR (25%), but lower than nested PCR (52.5%). The combination of both techniques, lysis-centrifugation and nested PCR, increased the overall sensitivity rate to 62.5%. These results have demonstrated that, globally, the nested PCR was more sensitive than both single PCR and lysis-centrifugation culture in detecting C. albicans in blood from immunecompetent rabbits with acute disseminated candidosis. PCR could be a useful complementary technique to traditional methods in the early diagnosis of candidemia.
Nexstar Farmaceutica (AMP), Madrid, Spain.
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A
Polanco
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E
Mellado
C
C
Castilla
J L
JL
Rodriguez-Tudela
D000818:Animals; D001769:Blood / Q000382:microbiology*; D015139:Blotting, Southern; D002176:Candida albicans / Q000235:genetics / Q000302:isolation & purification*; D002177:Candidiasis / Q000382:microbiology*; D002498:Centrifugation; D003470:Culture Media; D004271:DNA, Fungal / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D004195:Disease Models, Animal; D016469:Fungemia / Q000382:microbiology*; D008297:Male; D016133:Polymerase Chain Reaction; D012335:RNA, Ribosomal / Q000235:genetics; D011817:Rabbits; D012680:Sensitivity and Specificity
D003470:Culture Media; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012335:RNA, Ribosomal