title: Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems.
abstract: 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.
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Camila
authors.initials: C
authors.lastname: Ximenes
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Brandão
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Paula
authors.initials: P
authors.lastname: Oliveira
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Abraham
authors.initials: A
authors.lastname: Rocha
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Tamisa
authors.initials: T
authors.lastname: Rego
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Rafael
authors.initials: R
authors.lastname: Medeiros
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Ana
authors.initials: A
authors.lastname: Aguiar-Santos
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: João
authors.initials: J
authors.lastname: Ferraz
authors.affiliation: Departamento de Microbiologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Christian
authors.initials: C
authors.lastname: Reis
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Paulo
authors.initials: P
authors.lastname: Araujo
authors.affiliation: Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco, Recife, PE, Brasil.
authors.forename: Luiz
authors.initials: L
authors.lastname: Carvalho
authors.affiliation: Serviço de Referência Nacional em Filarioses, Departamento de Parasitologia, Centro de Pesquisas Aggeu Magalhães-Fiocruz, Recife, PE, Brasil.
authors.forename: Fabio L
authors.initials: FL
authors.lastname: Melo
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D000954:Antigens, Surface / Q000097:blood / Q000652:urine; D018509:DNA, Helminth / Q000302:isolation & purification*; D004605:Elephantiasis, Filarial / Q000097:blood / Q000175:diagnosis* / Q000652:urine; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D008842:Microfilariae / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D012680:Sensitivity and Specificity; D014958:Wuchereria bancrofti / Q000235:genetics / Q000302:isolation & purification*; D055815:Young Adult
chemical_list: D000954:Antigens, Surface; D018509:DNA, Helminth