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Diagnosis of invasive fungal infections using real‐time PCR assay in paediatric acute leukaemia induction
Author(s) -
Mandhaniya Sushil,
Iqbal Sobuhi,
Sharawat Surender Kumar,
Xess Immaculata,
Bakhshi Sameer
Publication year - 2012
Publication title -
mycoses
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.13
H-Index - 69
eISSN - 1439-0507
pISSN - 0933-7407
DOI - 10.1111/j.1439-0507.2011.02157.x
Subject(s) - voriconazole , medicine , aspergillosis , aspergillus , mucormycosis , amphotericin b , real time polymerase chain reaction , gastroenterology , pneumonia , candida infections , immunology , antifungal , biology , surgery , microbiology and biotechnology , dermatology , biochemistry , gene
Summary Invasive fungal infections (IFI) lead to morbidity and mortality in neutropenic patients and in allogenic stem cell transplantation. Serum‐based fungal detection assays have limitation of specificity or sensitivity. Studies on fungal DNA detection using real‐time PCR in childhood leukaemia are lacking. The aim of this study was to develop sensitive and specific diagnostic tools for IFI in paediatric acute leukaemia patients using real‐time PCR. Of 100 randomised paediatric acute leukaemia patients receiving antifungal prophylaxis with voriconazole/amphotericin B, single peripheral whole blood sample in EDTA was used for Pan‐AC real‐time PCR assay (detects nine Candida and six Aspergillus species) in patients who failed prophylaxis due to proven, probable, possible or suspected fungal infections. PCR results were retrospectively correlated with clinical profile. Real‐time PCR test was positive in 18/29 (62%) patients who failed prophylaxis. The only patient with proven IFI (mucormycosis), real‐time PCR assay was negative. Real‐time PCR was positive in 2/4 (50%) patients with possible and 16/24 (66.6%) suspected IFI and 5/10 (50%) patients with pneumonia. By applying method A/B, sensitivity and positive predictive value could not be commented due to unproven Aspergillus or Candida infections; specificity and negative predictive values (NPV) were 41% and 100% respectively; by method C (included episodes of possible IFI as true positive), sensitivity, specificity, PPV and NPV were 50%, 36%, 11% and 81% respectively. In those with suspected IFI, 8/24 (33.3%) were PCR negative and unnecessarily received empirical antifungal therapy (EAFT). Real‐time PCR is a practical, rapid, non‐invasive screening test for excluding IFI in paediatric leukaemia. The high NPV makes real‐time PCR a promising tool to use this prior to initiating EAFT in antibiotic‐resistant febrile neutropenic patients; this would avoid toxicity, cost and hospitalisation for EAFT (ClinicalTrials.gov identifier:NCT00624143).