Characterization of Cryptosporidium parvum in human and animal feces by single-tube nested polymerase chain reaction and restriction analysis.
A DNA isolation and purification method is described that produced DNA free of inhibitory substances in 148 of the 159 analyzed fecal samples. The polymerase chain reaction (PCR) product from a sensitive single-tube nested PCR that amplifies a part of an oocyst protein was used to characterize Cryptosporidium parvum genotypes by a simple restriction analysis. Genotype 1 was solely detected in human-derived oocysts, genotype 2 was present in both animal and human-derived oocysts. The ratio between both genotypes in humans in The Netherlands varied markedly between samples obtained during a period of augmented cases of cryptosporidiosis in the western part of the country and randomly selected samples from gastroenteritis patients. Sequence analysis of a 581-bp fragment from the nested PCR product revealed 12 nucleotide substitutions between the two genotypes. Sequences from isolates in each genotype group were identical.
Homan
W
W
Microbiological Laboratory for Health Protection, National Institute of Public Health and the Environment, Bilthoven, The Netherlands. wieger.homan@rivm.nl
van Gorkom
T
T
Kan
Y Y
YY
Hepener
J
J
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D016785:Cryptosporidium parvum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016054:DNA, Protozoan / Q000032:analysis*; D005243:Feces / Q000469:parasitology*; D006041:Goats; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep
D016054:DNA, Protozoan