title: Screening of invasive fungal infections by a real-time panfungal (pan-ACF) polymerase chain reaction assay in patients with haematological malignancy.
abstract: BACKGROUND
  Invasive fungal infection (IFI) is a fatal infection in haematology patients. There is an urgent need for reliable screening methods facilitating timely diagnosis and treatment. A real-time panfungal polymerase chain reaction (PCR) assay based on TaqMan technology targeting 18S ribosomal RNA gene was used to screen whole blood specimen obtained from series of Haematology malignancy patients for IFIs.
  
  
  METHODS
  The panfungal (Pan-ACF) assay was employed to investigate specimen from 133 patients in duplicate with suspected IFI. In addition twenty healthy subjects and twenty patients with bacterial infections were taken as control. The patients with suspected IFI were also diagnosed by conventional methods including direct microscopy, culture techniques and antigen detection (galactomannan antigen ELISA and latex agglutination for cryptococcal antigen). The results of molecular testing were evaluated in relation to the criteria proposed by the European Organization for Research and Treatment of Cancer and patients were classified as having proven and probable IFD.
  
  
  RESULTS
  Of 133 patients, 89 had proven, 18 had probable and 26 had possible IFI. One hundred four samples were reverse transcription-PCR positive. Of 89 proven cases, 84 were panfungal PCR positive. These 84 cases included 82 cases which revealed growth on fungal blood culture and two cases were negative on fungal blood culture. Of the 82 cases which revealed growth on culture: 74 grew Candida in culture, 3 grew Fusarium solani, 5 grew Aspergillus species on blood culture. The later five were also galactomannan antigen positive. The five specimen which were negative on panfungal PCR, two grew Trichosporon asahii, one grew Candida rugosa and two grew as Cryptococcus neoformans var. neoformans. Of the 18 probable cases, 18 were panfungal PCR positive. These were also galactomannan antigen positive. The sensitivity and specificity of panfungal PCR in proven cases were 94.3% and 95.2%, respectively. The positive and negative predictive values proven cases were 97.6% and 88.9%, respectively.
  
  
  CONCLUSIONS
  The panfungal (Pan-ACF) real-time PCR assay can detect common fungal genera and it may be used as an adjunct to conventional methods for screening of IFI.
authors.affiliation: Department of Microbiology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Malini Rajinder
authors.initials: MR
authors.lastname: Capoor
authors.affiliation: Department of Microbiology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Shikha
authors.initials: S
authors.lastname: Puri
authors.affiliation: Department of Medicine, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Hitesh
authors.initials: H
authors.lastname: Raheja
authors.affiliation: Department of Haematology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Ritin
authors.initials: R
authors.lastname: Mohindra
authors.affiliation: Department of Haematology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Dinesh Kumar
authors.initials: DK
authors.lastname: Gupta
authors.affiliation: Department of Medicine, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Pradeep Kumar
authors.initials: PK
authors.lastname: Verma
authors.affiliation: Independent Statistician, CHRD-SAS, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi, India.
authors.forename: Ranadeep
authors.initials: R
authors.lastname: Chowdhary
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D004271:DNA, Fungal / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005260:Female; D016469:Fungemia / Q000175:diagnosis*; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D019337:Hematologic Neoplasms / Q000150:complications*; D006801:Humans; D000072742:Invasive Fungal Infections / Q000175:diagnosis*; D008297:Male; D008403:Mass Screening / Q000379:methods*; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011446:Prospective Studies; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012337:RNA, Ribosomal, 18S