Development of a method for the detection of waterborne microsporidia

Rebecca M. Hoffman, Donna M. Wolk, Susan K. Spencer, Mark A. Borchardt

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Microsporidia are obligate intracellular pathogens capable of infecting humans. There is credible evidence to suggest that microsporidial infections may be transmitted through consumption of spores in contaminated water; however, methods to detect this pathogen have not been standardized and microsporidia occurrence studies have not been conducted. Concentration of spores by continuous flow centrifugation (CFC), purification using immunomagnetic separation (IMS), and detection by either microscopy or real-time polymerase chain reaction (PCR) were evaluated for detection of Encephalitozoon intestinalis spores in seeded water samples. Recovery efficiency of CFC using microscopic detection ranged from 38.7-75.5% in filtered tap water. Using an indirect IMS method, 78.8-90.2% of seeded spores were recovered in ultrapure water (18 MΩ); however, the lack of a specific monoclonal antibody and the presence of other particulates interfered with the IMS assay in some turbid samples. Despite low recovery efficiencies and the detectable presence of PCR inhibitors in each of the samples, a combination of CFC concentration, indirect IMS, and real-time PCR produced a positive test result in six of ten natural water samples (turbidity 0.1-28.9 NTU) at a seeding level of 50 spores/L.

Original languageEnglish (US)
Pages (from-to)312-318
Number of pages7
JournalJournal of Microbiological Methods
Volume70
Issue number2
DOIs
StatePublished - Aug 2007
Externally publishedYes

Keywords

  • Continuous flow centrifugation
  • Encephalitozoon
  • Enterocytozoon
  • Microsporidia

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

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