Genotyping human and bovine isolates of Cryptosporidium parvum by polymerase chain reaction-restriction fragment length polymorphism analysis of a repetitive DNA sequence.
In order to define transmission routes of cryptosporidiosis and develop markers that distinguish Cryptosporidium parvum isolates, we have identified 2 polymorphic restriction enzyme sites in a C. parvum repetitive DNA sequence. The target sequence was amplified by polymerase chain reaction from 100 to 500 oocysts and the amplified product was subjected to restriction enzyme digestion. Typing of 23 isolates showed that 10/10 calf isolates had the same profile. In contrast, 2 patterns were observed among human isolates: 7/13 displayed the calf profile, and 6/13 presented another pattern. The PCR-RFLP assay described here is a sensitive tool to distinguish C. parvum isolates.
Bonnin
A
A
Laboratoire de Parasitologie et Mycologie, Centre Hospitalier Universitaire de Dijon, France.
Fourmaux
M N
MN
Dubremetz
J F
JF
Nelson
R G
RG
Gobet
P
P
Harly
G
G
Buisson
M
M
Puygauthier-Toubas
D
D
Gabriel-Pospisil
G
G
Naciri
M
M
Camerlynck
P
P
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology / Q000635:transmission; D003001:Cloning, Molecular; D003457:Cryptosporidiosis / Q000469:parasitology / Q000635:transmission / Q000662:veterinary; D016785:Cryptosporidium parvum / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012091:Repetitive Sequences, Nucleic Acid; D015047:Zoonoses / Q000469:parasitology / Q000635:transmission
D017931:DNA Primers; D016054:DNA, Protozoan