Molecular identification and phylogenetic analysis of Wuchereria bancrofti from human blood samples in Egypt

Parasitol Res. 2017 Mar;116(3):963-970. doi: 10.1007/s00436-017-5372-7. Epub 2017 Jan 24.

Abstract

Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.

Keywords: Phylogenetic analysis; Repeated DNA sequence; Semi-nested PCR; Wuchereria bancrofti.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Animals
  • Base Sequence
  • Blood / parasitology*
  • Child
  • Child, Preschool
  • Culex / genetics
  • Egypt / epidemiology
  • Elephantiasis, Filarial / blood
  • Elephantiasis, Filarial / epidemiology
  • Elephantiasis, Filarial / parasitology*
  • Female
  • Humans
  • Infant
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Phylogeny*
  • Polymerase Chain Reaction
  • Sequence Alignment
  • Wuchereria bancrofti / classification
  • Wuchereria bancrofti / genetics*
  • Wuchereria bancrofti / isolation & purification
  • Young Adult