Detection of Hammondia heydorni-like organisms and their differentiation from Neospora caninum using random-amplified polymorphic DNA-polymerase chain reaction.
Neospora caninum and Hammondia heydorni are morphologically and phylogenetically related coccidians that are found in dogs. New diagnostic genetic loci, based on random-amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), were developed to aid in the detection of H. heydorni-like parasites and to discriminate them from N. caninum and other related coccidians of dogs. On the basis of the data obtained from 5 random decamers, H. heydorni (Manhattan-1) and N. caninum (NC1) were characterized by distinct banding patterns (similarity index = 0.068). High-stringency PCR assays were developed from the sequences of 2 cloned bands (GenBank BZ592549 and BZ592593), uniquely amplified from H. heydorni. Interestingly, using these primers, PCR amplification was achieved only from 2 of the 5 isolates presumed to represent H. heydorni. The same result was obtained from these 5 isolates using a recently described PCR assay directed to the H. heydorni internal transcribed spacer-1. It is concluded that H. heydorni and N. caninum are genetically distinct and that such tools may be useful for more detailed characterization of the diversity of related parasites occurring in dogs.
Sreekumar
C
C
Animal Parasitic Diseases Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Beltsville, Maryland 20705 , USA. kumar@anri.barc.usda.gov
Hill
D E
DE
Fournet
V M
VM
Rosenthal
B M
BM
Lindsay
D S
DS
Dubey
J P
JP
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary; D017124:Conserved Sequence; D016172:DNA Fingerprinting; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000032:analysis / Q000737:chemistry*; D004283:Dog Diseases / Q000469:parasitology; D004285:Dogs; D014644:Genetic Variation; D008969:Molecular Sequence Data; D018688:Neospora / Q000235:genetics / Q000302:isolation & purification*; D011110:Polymorphism, Genetic; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D021745:Sarcocystidae / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D016054:DNA, Protozoan