Detection and discrimination of Loa loa, Mansonella perstans and Wuchereria bancrofti by PCR-RFLP and nested-PCR of ribosomal DNA ITS1 region

作者：Jimenez Maribel^*; Miguel Gonzalez Luis; Carranza Cristina; Bailo Begona; Perez Ayala Ana; Muro Antonio; Luis Perez Arellano Jose; Garate Teresa

来源：Experimental Parasitology, 2011, 127(1): 282-286.

DOI：10.1016/j.exppara.2010.06.019

• Loa loa
• Mansonella perstans
• Wuchereria bancrofti
• ITS1
• PCR-RFLP
• Nested-PCR
• Differential detection

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 boa, 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.

• 出版日期2011-1