Cloning and characterization of an Onchocerca volvulus specific DNA sequence.
A cloned sequence, pOvs134, was isolated from a genomic library prepared from Onchocerca volvulus of savanna origin in the plasmid pUC9. pOvs134 hybridizes to all the geographic isolates of O. volvulus tested from both the New and the Old World, but not to the species Onchocerca gibsoni, Onchocerca gutturosa, Onchocerca ochengi, Onchocerca cervicalis, the filarial parasites Brugia malayi, or Dirofilaria immitis, nor to human or simuliid DNA. As little as 250 pg of DNA can be detected on a dot blot hybridization, suggesting that pOvs134 is sensitive enough to detect a single third stage larva. DNA sequence analysis of the inserted DNA of pOvs134 revealed that it consisted of twelve examples of a 149-bp repeat. The sequence of this repeat is strikingly similar to that of two O. volvulus genomic clones previously described, one of which has been reported to be specific for forest form O. volvulus, and one of which hybridizes to genomic DNA of several species of Onchocerca. These results suggest that the 149-bp repeat sequence is highly repeated in the genome of O. volvulus, and that variants of this repeat with different specificities exist.
Meredith
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Department of Tropical Public Health, Boston, MA 02115.
Unnasch
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Karam
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Piessens
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Wirth
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D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005784:Gene Amplification; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009854:Onchocerca / Q000235:genetics*; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D012843:Simuliidae / Q000235:genetics
D004247:DNA