Novel Multiplex Polymerase Chain Reaction Approach for Detection of Four Human Infective Cryptosporidium Isolates: Cryptosporidium Parvum, types H and C, Cryptosporidium Canis, and Cryptosporidium Felis in Fecal and Soil Samples

A nested multiplex polymerase chain reaction (PCR) approach was adopted for the simultaneous detection of 4 human infective genotypes of the protozoan parasite Cryptosporidium. Specific PCR primers were designed for the heat shock protein 70 gene of 2 genotypes of Cryptosporidium parvum (human and b...

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Bibliographic Details
Published in:Journal of veterinary diagnostic investigation 2003-05, Vol.15 (3), p.262-267
Main Authors: Lindergard, Gabriella, Nydam, Daryl V, Wade, Susan E, Schaaf, Stephanie L, Mohammed, Hussni O
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cats
Cattle
cross reaction
Cryptosporidium - classification
Cryptosporidium - genetics
Cryptosporidium - isolation & purification
Cryptosporidium parvum
detection limit
DNA
DNA primers
Dogs
feces
Feces - microbiology
Fundamental and applied biological sciences. Psychology
genes
Genotype
heat shock proteins
Humans
Microbiology
oocysts
parasites
polymerase chain reaction
Polymerase Chain Reaction - methods
Sensitivity and Specificity
soil
Soil Microbiology
soil sampling
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Summary:A nested multiplex polymerase chain reaction (PCR) approach was adopted for the simultaneous detection of 4 human infective genotypes of the protozoan parasite Cryptosporidium. Specific PCR primers were designed for the heat shock protein 70 gene of 2 genotypes of Cryptosporidium parvum (human and bovine types), Cryptosporidium canis, and Cryptosporidium felis. These 4 genotypes have all been found in human fecal samples. The primers amplified DNA fragments of specific sizes, each representing a unique genotype. The limit of detection of the method was found to vary between 10 and 100 oocysts per 1 ml fecal material. There appeared to be no cross-reactivity with other organisms commonly present in feces and soil, and the approach has a high specificity. The rapid identification of various human infective Cryptosporidium isolates is a part of the authors' long-term aim of determining the routes of infection with oocysts and thereby increase their epidemiological understanding of Cryptosporidium infection in humans and animals.
ISSN:1040-6387
1943-4936
DOI:10.1177/104063870301500307