Brugia malayi: Whole genome amplification for genomic characterization of filarial parasites

• Published in: Experimental parasitology 2008-06, Vol.119 (2), p.256-263
• Main Authors: McNulty, Samantha N., Weil, Gary J., Heinz, Michael, Crosby, Seth D., Fischer, Peter U.
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Brugia malayi; Brugia malayi - genetics; Cycle threshold ( Ct); DNA Primers - chemistry; DNA, Helminth - chemistry; DNA, Helminth - isolation & purification; DNA, Ribosomal - chemistry; Electrophoresis, Agar Gel; Filarioidea; Fundamental and applied biological sciences. Psychology; Gene Amplification; Genome, Helminth - genetics; Gerbillinae; Helminth Proteins - genetics; Life cycle. Host-agent relationship. Pathogenesis; Microarray; Multiple displacement amplification (MDA); Nematoda; Oligonucleotide Array Sequence Analysis; Oligonucleotide microarray; Polymerase Chain Reaction - methods; Protozoa; Real-time PCR; RNA, Ribosomal, 18S - genetics; Whole genome amplification (WGA)
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2008.02.003

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.