title: Detection and discrimination of Loa loa, Mansonella perstans and Wuchereria bancrofti by PCR-RFLP and nested-PCR of ribosomal DNA ITS1 region.
abstract: The ribosomal deoxyribonucleic acid (DNA) internal transcribed spacer region (ITS1) of two filarial nematodes, Loa loa and Mansonella perstans, was amplified and further sequenced to develop an species-specific polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol for the differentiation of both species from Wuchereria bancrofti, three filarial nematodes with blood circulating microfilariae. The ITS1-PCR product digested with the restriction endonuclease Ase I generated an specific diagnostic pattern for each of the three species. Moreover, three new specific nested-PCRs, targeting the ITS1 region, for differential detection of L. loa, M. perstans and W. bancrofti were developed and used when the ITS1-PCR products were insufficient for the Ase I enzymatic digestion. These filarial species-specific molecular protocols were evaluated in forty blood samples from African adult immigrants attending in the Hospital Insular of Gran Canaria, Canarias, Spain.
authors.affiliation: Servicio de Parasitología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain. mjimenez@isciii.es
authors.forename: Maribel
authors.initials: M
authors.lastname: Jiménez
authors.forename: Luis Miguel
authors.initials: LM
authors.lastname: González
authors.forename: Cristina
authors.initials: C
authors.lastname: Carranza
authors.forename: Begoña
authors.initials: B
authors.lastname: Bailo
authors.forename: Ana
authors.initials: A
authors.lastname: Pérez-Ayala
authors.forename: Antonio
authors.initials: A
authors.lastname: Muro
authors.forename: José Luis
authors.initials: JL
authors.lastname: Pérez-Arellano
authors.forename: Teresa
authors.initials: T
authors.lastname: Gárate
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000097:blood / Q000737:chemistry; D004275:DNA, Ribosomal / Q000737:chemistry; D003937:Diagnosis, Differential; D004605:Elephantiasis, Filarial / Q000175:diagnosis* / Q000469:parasitology; D006801:Humans; D008117:Loa / Q000235:genetics / Q000302:isolation & purification*; D008118:Loiasis / Q000175:diagnosis* / Q000469:parasitology; D008366:Mansonella / Q000235:genetics / Q000302:isolation & purification*; D008368:Mansonelliasis / Q000175:diagnosis* / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D018510:RNA, Helminth / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D016415:Sequence Alignment; D013045:Species Specificity; D014958:Wuchereria bancrofti / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D018509:DNA, Helminth; D004275:DNA, Ribosomal; D018510:RNA, Helminth; D012335:RNA, Ribosomal