Dirofilaria repens: cloning and characterization of a repeated DNA sequence for the diagnosis of dirofilariasis in dogs, Canis familiaris.
A highly repetitive DNA element from the genome of the filarial nematode Dirofilaria repens has been cloned and sequenced. The 176-base pair repeating units are arranged in direct tandem and are clustered in the parasite genome. All repeats appear to belong to a single family although some elements have diverged significantly. The repeats are present in about 15,000 copies and constitute approximately 3.0% of the parasite genome. The cloned repetitive sequence hybridized only to D. repens DNA and was sensitive enough to detect 250 to 500 pg of D. repens DNA, a single microfilariae in infected blood samples, and a single third stage larvae in mosquitoes. The high specificity and sensitivity of the cloned fragment makes it ideal as a diagnostic probe for detecting D. repens in both the host and the vector.
Chandrasekharan
N V
NV
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Colombo, Sri Lanka.
Karunanayake
E H
EH
Franzen
L
L
Abeyewickreme
W
W
Pettersson
U
U
D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004182:Dirofilaria / Q000235:genetics* / Q000302:isolation & purification; D004184:Dirofilariasis / Q000175:diagnosis*; D004283:Dog Diseases / Q000175:diagnosis*; D004285:Dogs; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D015342:DNA Probes; D004247:DNA