Elsevier

Molecular Diagnosis

Volume 4, Issue 1, March 1999, Pages 57-64
Molecular Diagnosis

Original Research
Fast and reliable extraction of protozoan parasite DNA from fecal specimens2

https://doi.org/10.1016/S1084-8592(99)80050-2Get rights and content

Background:

Polymerase chain reaction (PCR) detection of intestinal protozoa in fecal specimens is hampered by poor recovery of DNA and by the presence of PCR inhibitors. In this study we describe a novel method for DNA extraction from such specimens containing spores and oocysts of Enterocytozoon bieneusi and Cryptosporidium parvum, respectively.

Methods and Results:

Extraction was done using commercial kits modified to maximize the recovery and purity of extracted DNA. In comparison with a procedure we previously reported, we estimate that this method may increase the sensitivity of parasite DNA detection in fecal specimens up to tenfold. An additional advantage of this method is that up to 12 samples may be processed simultaneously within 2 hours.

Conclusions:

By using this method, we were able to increase reproducibility of PCR amplification on fecal specimens and significantly reduce the hands-on time required to process the samples.

References (28)

  • AshL et al.

    Collection and preservation of feces. Parasites: A guide to laboratory procedures and identification

  • GarciaLS et al.

    Detection of microsporidial spores in fecal specimens from patients diagnosed with cryptosporidiosis

    J Clin Microbiol

    (1994)
  • SchwartzDA

    Emerging and reemerging infections Progress and challenges in the subspecialty of infectious disease pathology

    Arch Pathol Lab Med

    (1997)
  • SchwartzDA et al.

    Microsporidia

  • WeberR et al.

    Human microsporidial infections

    Clin Microbiol Rev

    (1994)
  • da SilvaAJ et al.

    Diagnosis of Enterocytozoon bieneusi (Microsporidia) infections by polymerase chain reaction in stool samples using primers based on the region coding for small-subunit ribosomal RNA

    Arch Pathol Lab Med

    (1997)
  • JohnsonDW et al.

    Development of a PCR protocol for sensitive detection of Cryptosporidium parvum oocysts in water samples

    Appl Environ Microbiol

    (1995)
  • TalalAH et al.

    Detection of Enterocytozoon bieneusi in fecal specimens by polymerase chain reaction

    Clin Infect Dis

    (1998)
  • Bornay-LlinaresFJ et al.

    Immunologic, microscopic, and molecular evidence of Encephalitozoon intestinalis (syn. Septata intestinalis) infection in mammals other than humans

    J Infect Dis

    (1998)
  • ChomczynskiP et al.

    DNAzol. A reagent for the rapid isolation of genomic DNA

    BioTechniques

    (1997)
  • OmbrouckC et al.

    Specific PCR assay for direct detection of intestinal microsporidia Enterocytozoon bieneusi and Encephalitozoon intestinalis in fecal specimens from human immunodeficiency virus-infected patients

    J Clin Microbiol

    (1997)
  • VelasquezJN et al.

    Detection of the microsporidian parasite Enterocytozoon bieneusi in specimens from patients with AIDS by PCR

    J Clin Microbiol

    (1996)
  • WeberR et al.

    Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates

    N Engl J Med

    (1992)
  • MouraH et al.

    A new and improved “quick-hot gram-chromotrpe” technique that differentially stains microsporidian spores in clinical samples, including paraffin-embedded tissue sections

    Arch Pathol Lab Med

    (1997)
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