Detection of Fungal DNA in Human Body Fluids and Tissues during a Multistate Outbreak of Fungal Meningitis and Other Infections
- 1 May 2013
- journal article
- research article
- Published by American Society for Microbiology in Eukaryotic Cell
- Vol. 12 (5), 677-683
- https://doi.org/10.1128/ec.00046-13
Abstract
Exserohilum rostratum was the major cause of an outbreak of fungal infections linked to injections of contaminated methylprednisolone acetate. Because almost 14,000 persons were exposed to product that was possibly contaminated with multiple fungal pathogens, there was unprecedented need for a rapid throughput diagnostic test that could detect both E. rostratum and other unusual agents of fungal infection. Here we report development of a novel PCR test that allowed for rapid and specific detection of fungal DNA in cerebrospinal fluid (CSF), other body fluids and tissues of infected individuals. The test relied on direct purification of free-circulating fungal DNA from fluids and subsequent PCR amplification and sequencing. Using this method, we detected Exserohilum rostratum DNA in 123 samples from 114 case-patients (28% of 413 case-patients for whom 627 samples were available), and Cladosporium DNA in one sample from one case-patient. PCR with novel Exserohilum -specific ITS-2 region primers detected 25 case-patients with samples that were negative using broad-range ITS primers. Compared to fungal culture, this molecular test was more sensitive: of 139 case-patients with an identical specimen tested by culture and PCR, E. rostratum was recovered in culture from 19 (14%), but detected by PCR in 41 (29%), showing a diagnostic sensitivity of 29% for PCR compared to 14% for culture in this patient group. The ability to rapidly confirm the etiologic role of E. rostratum in these infections provided an important contribution in the public health response to this outbreak.Keywords
This publication has 32 references indexed in Scilit:
- A Real-Time PCR Assay for Rapid Detection and Quantification of Exserohilum rostratum, a Causative Pathogen of Fungal Meningitis Associated with Injection of Contaminated MethylprednisoloneJournal of Clinical Microbiology, 2013
- Diverse Lifestyles and Strategies of Plant Pathogenesis Encoded in the Genomes of Eighteen Dothideomycetes FungiPLoS Pathogens, 2012
- The Index Case for the Fungal Meningitis Outbreak in the United StatesNew England Journal of Medicine, 2012
- Molecular Identification andIn VitroResponse to Antifungal Drugs of Clinical Isolates of ExserohilumAntimicrobial Agents and Chemotherapy, 2012
- Therapy with antifungals decreases the diagnostic performance of PCR for diagnosing invasive aspergillosis in bronchoalveolar lavage samples of patients with haematological malignanciesJournal of Antimicrobial Chemotherapy, 2012
- Critical Stages of Extracting DNA from Aspergillus fumigatus in Whole-Blood SpecimensJournal of Clinical Microbiology, 2010
- Validation of Laboratory-Developed Molecular Assays for Infectious DiseasesClinical Microbiology Reviews, 2010
- Aspergillus PCR: One Step Closer to StandardizationJournal of Clinical Microbiology, 2010
- Detection of Aspergillus DNA in Cerebrospinal Fluid from Patients with Cerebral Aspergillosis by a Nested PCR AssayJournal of Clinical Microbiology, 2006
- Exserohilum: an emerging human pathogenEuropean Journal of Clinical Microbiology & Infectious Diseases, 2006