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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Real-time PCR detection and speciation of Cryptosporidium infection using Scorpion probes
Journal of Medical Microbiology, Volume 55, No. 9, Year 2006
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Description
At least eight species of Cryptosporidium can cause human infection and disease. A real-time PCR (qPCR) assay based on the 18S rRNA gene and utilizing a Scorpion probe was developed to detect all human-pathogenic Cryptosporidium without the usual need for nested amplification. Sensitivity of detection in stool samples was highest using a glass bead-based DNA extraction method (under 103 oocysts per stool sample). The assay was validated against 123 human stool specimens from Bangladesh and Tanzania, exhibited a sensitivity and specificity of >91% versus microscopy, and detected an additional eight microscopy-negative infections. Cryptosporidium parvum-specific and Cryptosporidium meleagridis-specific Scorpion qPCR assays that provided 100% accurate speciation compared with VspI RFLP analysis and sequencing were developed subsequently. These Scorpion probe qPCR assays are simpler to perform than existing nested PCR and RFLP methods for diagnosis and epidemiological investigation of cryptosporidiosis. © 2006 SGM.
Authors & Co-Authors
Stroup, Suzanne E.
United States, Charlottesville
University of Virginia
Roy, Shantanu
Bangladesh, Dhaka
Icddr, b Centre for Health and Population Research
Mchele, John P D
Tanzania, Moshi
Kilimanjaro Christian Medical Centre
Maro, Venance Phillip
Tanzania, Moshi
Kilimanjaro Christian Medical Centre
Ntabaguzi, Simon Angelo
Tanzania, Moshi
Kilimanjaro Christian Medical Centre
Siddique, Abdullah B.
Bangladesh, Dhaka
Icddr, b Centre for Health and Population Research
Kang, Gagandeep
India, Vellore
Christian Medical College, Vellore
Guerrant, Richard Littleton
United States, Charlottesville
University of Virginia
Kirkpatrick, Beth D.
United States, Burlington
University of Vermont Larner College of Medicine
Fayer, Ronald
United States, Beltsville
Usda Ars Beltsville Agricultural Research Center
Herbein, Joel F.
United States, Blacksburg
Techlab, Inc.
Ward, Honorine D.
United States, Medford
Tufts University
Haque, Rashidul
Bangladesh, Dhaka
Icddr, b Centre for Health and Population Research
Houpt, Eric R.
United States, Charlottesville
University of Virginia
Statistics
Citations: 49
Authors: 14
Affiliations: 8
Identifiers
Doi:
10.1099/jmm.0.46678-0
ISSN:
00222615
Research Areas
Genetics And Genomics
Study Locations
Tanzania