4 Scopus citations

Abstract

Genetic characterization of field isolates and clinical specimens of filarial nematodes is often limited by a shortage of DNA; therefore, we evaluated a multiple displacement amplification (MDA) based whole genome amplification method. The quality of amplified DNA was examined by conventional PCR, real-time PCR, and DNA hybridization. MDA of 5.0 ng of adult Brugia malayi DNA and one-fifteenth of the DNA isolated from a single microfilaria resulted in 6.3 and 4.2 μg of amplified DNA, respectively. Amplified DNA was equivalent to native genomic DNA for hybridization to B. malayi BAC library clones or to an oligonucleotide microarray with approximately 18,000 filarial DNA sequences. MDA is useful for whole genome amplification of filarial DNA from very small amounts of starting material. This technology will permit detailed studies of genetic diversity that were not previously feasible.

Original languageEnglish
Pages (from-to)256-263
Number of pages8
JournalExperimental Parasitology
Volume119
Issue number2
DOIs
StatePublished - Jun 1 2008

Keywords

  • Brugia malayi
  • Cycle threshold (C)
  • Microarray
  • Multiple displacement amplification (MDA)
  • Oligonucleotide microarray
  • Real-time PCR
  • Whole genome amplification (WGA)

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