Brugia malayi: whole genome amplification for genomic characterization of filarial parasites.
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 microg 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.
McNulty
Samantha N
SN
Internal Medicine/Infectious Diseases Division, Washington University School of Medicine, Campus Mailbox 8051, 660 S. Euclid Avenue, St. Louis, MO 63110, USA.
Weil
Gary J
GJ
Heinz
Michael
M
Crosby
Seth D
SD
Fischer
Peter U
PU
D000818:Animals; D017178:Brugia malayi / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D049751:Genome, Helminth / Q000235:genetics*; D005849:Gerbillinae; D015801:Helminth Proteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D012337:RNA, Ribosomal, 18S / Q000235:genetics
D017931:DNA Primers; D018509:DNA, Helminth; D004275:DNA, Ribosomal; D015801:Helminth Proteins; D012337:RNA, Ribosomal, 18S