1921
Volume 54, Issue 4
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645
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Abstract

Abstract

To identify DNA sequences that could be used as the basis for a simple and rapid parasite detection assay, a genomic library of was constructed and screened for highly repeated DNA. The repeat found with the highest copy number was 195 basepairs (bps) long, 77% AT, and 300 copies per haploid genome. This sequence was designated the I repeat because it has a unique recognition site for that restriction endonuclease in all or most of the repeat copies. The I repeat DNA family is dispersed, genus-specific, and exists in all of the different geographic isolates of tested. Based on DNA sequence analysis of this repeat, we have developed an assay to detect very small quantities of DNA using the polymerase chain reaction (PCR). With this PCR assay, the I repeat was detected in as little as 1 pg of genomic DNA (about 1% of the DNA in one microfilaria) added to 100 µl of human blood. The PCR assay also amplified I repeat DNA from geographic isolates of from around the world but not from other species of filariae or from human or mosquito DNA. Microfilaria-positive human blood samples collected in Mauke, Cook Islands were shown to be I PCR-positive, while microfilaria-negative samples were PCR-negative. The specificity and sensitivity of the I PCR assay indicates that this approach has significant potential for improved screening of large human populations for active infection.

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/content/journals/10.4269/ajtmh.1996.54.357
1996-04-01
2017-11-25
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