Improved detection of Dirofilaria repens DNA by direct polymerase chain reaction.
Diagnosis of human infection by Dirofilaria repens, depends mainly on microscopic evaluation of tissue cross-sections and the macroscopic characteristics of the worm. Tissue degeneration and/or poor specimen preparation practices however, often render many cases of subcutaneous dirofilariasis elusive to such morphological diagnostic approaches. The early PCR protocols, developed to satisfy these complex diagnostic needs, failed to amplify dirofilariae DNA from formalin preserved material. To overcome these difficulties, we developed an improved PCR protocol using a set of primers designed to amplify a rather stable, highly repetitive D. repens-specific genomic DNA target. We report the performance of this protocol with a large variety of dirofilariae infected DNA specimens, including those extracted from formalin preserved biological material for up to 20 days. Our findings support its potential application to routine clinical diagnosis.
Department of Parasitology, Entomology and Tropical Diseases, National School of Public Health, Athens, Attki, Greece.
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N
Vakalis
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G
Spanakos
E
E
Patsoula
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NC
Vamvakopoulos
D000818:Animals; D017931:DNA Primers; D018509:DNA, Helminth / Q000032:analysis*; D004182:Dirofilaria / Q000235:genetics / Q000302:isolation & purification*; D004184:Dirofilariasis / Q000175:diagnosis* / Q000469:parasitology; D004283:Dog Diseases / Q000469:parasitology; D004285:Dogs; D005557:Formaldehyde; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D013048:Specimen Handling
D017931:DNA Primers; D018509:DNA, Helminth; D005557:Formaldehyde