Duplex Real-Time PCR Assay Distinguishes Aedes aegypti From Ae. albopictus (Diptera: Culicidae) Using DNA From Sonicated First-Instar Larvae

Details

• Alternative Title:
  J Med Entomol
• Personal Author:
  Kothera, Linda ; Byrd, Brian ; Savage, Harry M.
• Description:
  Aedes aegypti (L.) and Ae. albopictus (Skuse) are important arbovirus vectors in the United States, and the recent emergence of Zika virus disease as a public health concern in the Americas has reinforced a need for tools to rapidly distinguish between these species in collections made by vector control agencies. We developed a duplex real-time PCR assay that detects both species and does not cross-amplify in any of the other seven Aedes species tested. The lower limit of detection for our assay is equivalent to ∼0.03 of a first-instar larva in a 60-µl sample (0.016 ng of DNA per real-time PCR reaction). The assay was sensitive and specific in mixtures of both species that reflected up to a 2,000-fold difference in DNA concentration. In addition, we developed a simple protocol to extract DNA from sonicated first-instar larvae, and used that DNA to test the assay. Because it uses real-time PCR, the assay saves time by not requiring a separate visualization step. This assay can reduce the time needed for vector control agencies to make species identifications, and thus inform decisions about surveillance and control.
• Subjects:
  Aedes Aedes Aegypti Aedes Albopictus Animals Article Base Sequence DNA Larva Real-time PCR Real-Time Polymerase Chain Reaction Species Identification Assay Zika

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• Source:
  J Med Entomol. 54(6):1567-1572
• Pubmed ID:
  28981691
• Pubmed Central ID:
  PMC6437760
• Document Type:
  Journal Article
• Funding:
  CC999999/Intramural CDC HHS/United States
• Volume:
  54
• Issue:
  6
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:55c8fe5b9937665591c02d76f12bd474c015c210a9f558cd72cef79554491781
• Download URL:
  https://stacks.cdc.gov/view/cdc/77025/cdc_77025_DS1.pdf
• File Type:
  [PDF - 771.08 KB ]