title: Standardization of fungal polymerase chain reaction for the early diagnosis of invasive fungal infection.
abstract: BACKGROUND
  An early initiation of antifungal therapy in invasive fungal infections (IFIs) is critical in reducing the high mortality rate. Current diagnosis of fungal infection relies on microscopy, culture, antigen, antibody specific tests and histological diagnosis. However, these tests either lack sensitivity or specificity. There is thus the need for a rapid, specific and accurate diagnostic method.
  
  
  OBJECTIVE
  The aim of our study was to establish PCR for the rapid detection of Candida and Aspergillus species in clinical specimens with improved sensitivity and specificity.
  
  
  METHODS
  A total of 71 proven cases of IFI (confirmed by culture) were collected. A total of 15 healthy, 15 patients suffering from bacterial sepsis and 15 patients with HIV, HBV viral infections were included as controls. Clinical specimens were subjected to a standardized nested amplification to produce Round I (504 bp) and Round II (150 bp) amplicons. Restriction digestion was performed on these products for further identification.
  
  
  RESULTS
  Analytical sensitivity was determined using 10⁶-10 CFU/ml of cell suspension. The lower detection limit of the assay was 10 CFU/ml of blood. This test was 100% sensitive and specific with a positive predictive value of 100% and a negative predictive value of 96.7%.
  
  
  CONCLUSIONS
  The assay was found to be effective for the rapid detection of Candida and Aspergillus in clinical specimens.
authors: Department of Research, PD Hinduja National Hospital and Medical Research Centre, Veer Savarkar Marg, Mahim, Mumbai 400 016, Maharashtra, India.
authors: P
authors: P
authors: Deshpande
authors: A
authors: A
authors: Shetty
authors: A
authors: A
authors: Mehta
authors: F
authors: F
authors: Kapadia
authors: A
authors: A
authors: Hedge
authors: R
authors: R
authors: Soman
authors: C
authors: C
authors: Rodrigues
mesh_terms: D001228:Aspergillosis / Q000175:diagnosis*; D001230:Aspergillus / Q000235:genetics / Q000302:isolation & purification; D002175:Candida / Q000235:genetics / Q000302:isolation & purification; D002177:Candidiasis / Q000175:diagnosis*; D042241:Early Diagnosis; D016469:Fungemia / Q000175:diagnosis*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards*; D009172:Mycology / Q000379:methods / Q000592:standards*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D012680:Sensitivity and Specificity