Differentiation of Cryptosporidium parvum, C. muris, and C. baileyi by pcr-rflp analysis of the 18s rrna gene

We have developed a combined polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay for the detection and differentiation of Cryptosporidium parvum, Cryptosporidium muris, and Cryptosporidium baileyi. The assay utilizes PCR to amplify an approximately 1750 basepair pro...

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Bibliographic Details
Published in:Veterinary parasitology 1996-03, Vol.62 (1), p.1-7
Main Authors: Xigang Leng, Mosier, Derek A., Oberst, Richard D.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Cryptosporidium - classification
Cryptosporidium - genetics
Cryptosporidium - isolation & purification
Cryptosporidium parvum - classification
Cryptosporidium parvum - genetics
Cryptosporidium parvum - isolation & purification
Cryptosporidium spp
Deoxyribonucleases, Type II Site-Specific
Diagnosis-Protozoa
DNA Primers
Molecular Sequence Data
Polymerase chain reaction
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Restriction fragment length polymorphism assay
Restriction Mapping
RNA, Ribosomal, 18S - genetics
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Summary:We have developed a combined polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay for the detection and differentiation of Cryptosporidium parvum, Cryptosporidium muris, and Cryptosporidium baileyi. The assay utilizes PCR to amplify an approximately 1750 basepair product of the Cryptosporidium 18S rRNA gene. Following double digestion with restriction endonucleases DraI and VspI, this PCR product yields DNA fragments that can be resolved electrophoretically into RFLP profiles that are distinct for C. parvum, C. muris, and C. baileyi. Previous PCR-restriction analyses could not simultaneously differentiate all three species. Future application of this technique could include predicting the disease-causing potential of oocyst-contaminated environmental specimens and helping to determine the source of oocyst contamination.
ISSN:0304-4017
1873-2550
DOI:10.1016/0304-4017(95)00863-2