1. Detection of Naegleria fowleri, Acanthamoeba spp, and Balamuthia mandrillaris in Formalin-Fixed, Paraffin-Embedded Tissues by Real-Time Multiplex Polymerase Chain Reaction
- Author
-
Bobbi S. Pritt, Lynne M. Sloan, and Andrew P. Norgan
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,Tissue Fixation ,Formalin fixed paraffin embedded ,030106 microbiology ,Acanthamoeba ,Real-Time Polymerase Chain Reaction ,Sensitivity and Specificity ,Balamuthia mandrillaris ,law.invention ,03 medical and health sciences ,law ,Formaldehyde ,Multiplex polymerase chain reaction ,medicine ,Humans ,Multiplex ,Naegleria fowleri ,Polymerase chain reaction ,Paraffin Embedding ,biology ,Amebiasis ,General Medicine ,biology.organism_classification ,030104 developmental biology ,Tissue sections ,Multiplex Polymerase Chain Reaction - Abstract
Objectives Pathogenic free-living amebae (FLAs) cause skin, ocular, and central nervous system (CNS) infections with significant morbidity and mortality. Diagnosis of FLA infections by pathologic examination of tissue sections can be aided using molecular assays. This study investigated the performance characteristics of a multiplex real-time polymerase chain reaction (PCR) assay (FLA-PCR) for detection and differentiation of FLAs in clinical specimens. Methods FLA-PCR was performed on 39 human specimens comprising one cutaneous, 14 corneal, and 24 CNS formalin-fixed, paraffin-embedded (FFPE) tissues with a histopathologic diagnosis of FLA infection and four CNS FFPE tissues with inflammation but no evidence of FLAs. In addition, clinical specificity and assay limit of detection were determined. Results FLA detection sensitivities ranged from 79% to 84% in FFPE tissues. No cross-reactivity was observed. Conclusions While sensitivity is limited, FLA-PCR assay may serve as a useful adjunct for detection or confirmation of FLA infections in FFPE tissues.
- Published
- 2019