Detection of Blastocystis from stool samples using real-time PCR (original) (raw)

Abstract

We developed a real-time LC PCR assay to detect a 152 bp sequence in an uncharacterized region of the Blastocystis genome. The described assay detected 11 of 11 ATCC strains of Blastocystis from subtypes 1, 3, and 4. Three of three stool samples from Oregon and California military personnel that were negative for Blastocystis by an ova and parasite test as well as a conventional PCR assay were positive for Blastocystis using our real-time LC PCR assay. Diagnosis of Blastocystis infections using this sensitive method, including DNA extraction and real-time PCR, only requires 3 h. The lower limit of detection for Blastocystis in stool using the real-time LC PCR assay was calculated to be 760 cells of Blastocystis per 100 mg of stool, an estimated 760 parasites per reaction. The assay did not cross-react with Ruminococcus hansenii, Anarococcus hydrogenalis, Bifidobacterium adolescentis, Fusobacterium prausnitzii, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, or Lactobacillus acidophilus. Because of the ease of use, sensitivity, specificity, and increase in Blastocystis infections in the USA we believe this assay has the potential to be useful as a clinical diagnosis tool of Blastocystis infection.

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Acknowledgments

The work reported herein was performed under United States Air Force Surgeon General-approved Clinical Investigation No. FDG20070009N, FDG20070010N. The experiments comply with the current laws of the country in which they were performed. The views expressed in this material are those of the authors, and do not reflect the official policy or position of the U.S. Government, the Department of Defense, or the Department of the Air Force.

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Authors and Affiliations

1. Clinical Investigation Facility, David Grant USAF Medical Center, 101 Bodin Circle, Travis AFB, CA, 94535, USA
   Morris Saffold Jones II, Robert D. Ganac, Greg Hiser, N. Ryan Hudson & Andy Le
2. Environmental and Forest Biology, SUNY College of Environmental Science & Forestry 246 Illick, 1 Forestry Dr., Syracuse, NY, 13210, USA
   Christopher M. Whipps

Authors

1. Morris Saffold Jones II
2. Robert D. Ganac
3. Greg Hiser
4. N. Ryan Hudson
5. Andy Le
6. Christopher M. Whipps

Corresponding author

Correspondence toMorris Saffold Jones II.

Additional information

The work reported herein was performed under United States Air Force Surgeon General-approved Clinical Investigation No. FDG20070009N, FDG20070010N.

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Jones II, M.S., Ganac, R.D., Hiser, G. et al. Detection of Blastocystis from stool samples using real-time PCR.Parasitol Res 103, 551–557 (2008). https://doi.org/10.1007/s00436-008-1006-4

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• Received: 14 April 2008
• Accepted: 16 April 2008
• Published: 17 May 2008
• Issue date: August 2008
• DOI: https://doi.org/10.1007/s00436-008-1006-4

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