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Mem. Inst. Oswaldo Cruz ; 109(8): 978-983, 12/2014. graf
Article Dans Anglais | LILACS | ID: lil-732610

Résumé

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.


Sujets)
Adolescent , Adulte , Animaux , Femelle , Humains , Mâle , Adulte d'âge moyen , Jeune adulte , ADN des helminthes/isolement et purification , Filariose lymphatique/diagnostic , Microfilaria/isolement et purification , Wuchereria bancrofti/isolement et purification , Antigènes de surface/sang , Antigènes de surface/urine , Filariose lymphatique/sang , Filariose lymphatique/urine , Limite de détection , Microfilaria/génétique , Réaction de polymérisation en chaîne/méthodes , Réaction de polymérisation en chaîne/normes , Sensibilité et spécificité , Wuchereria bancrofti/génétique
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