Species-Specific Sequence in the Repeat 3 Region of the Gene Encoding a Putative Loa loa Allergen: a Diagnostic Tool for Occult Loiasis

Fousseyni S. ToureCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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Thomas G. EgwangCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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Goetz WahlCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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Pascal MilletCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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Odile BainCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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Alain J. GeorgesCentre International de Recherches Medicales de Franceville, Laboratoire de Biologie Parasitaire Museum National d'Histoire Naturelle, Franceville, Gabon

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A polymerase chain reaction (PCR)-based method to detect Loa loa DNA in the blood lysate of infected individuals is described. A set of primers was designed to amplify the repeat 3 sequence (15r3) of the gene encoding a putative L. loa allergen. The qualitative PCR was carried out using blood lysates from subjects from an L. loa-endemic area of Gabon where loiasis exists sympatrically with Mansonella perstans, and from individuals from a loiasis-free area in Togo infected concomitantly with M. perstans and Onchocerca volvulus. No specific amplification was observed after ethidium bromide staining of a gel containing M. perstans and O. volvulus control samples. In contrast, a 396-basepair (bp) DNA was detected in all L. loa microfilaremic individuals and in seven of the 20 L. loa amicrofilaremic subjects diagnosed by leukoconcentration. Qualitative Southern blots carried out at high stringency (65°C) using 15r3 oligonucleotide probe revealed hybridization only with L. loa samples (5 of 5 microfilaremic individuals and 15 of 20 amicrofilaremic individuals), confirming the results obtained with ethidium bromide staining of PCR products. We conclude that this 396-bp sequence could be used as a species-specific diagnostic tool for occult loiasis in an endemic area with concurrent filarial infections.

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