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Member species of the Anopheles gambiae complex can be misidentified as Anopheles leesoni

Authors
  • Dahan-Moss, Yael1, 2
  • Hendershot, Allison3
  • Dhoogra, Minishca1, 2
  • Julius, Henry1
  • Zawada, Jacek1, 2
  • Kaiser, Maria1, 2
  • Lobo, Neil F.3
  • Brooke, Basil D.1, 2
  • Koekemoer, Lizette L.1, 2
  • 1 Centre for Emerging Zoonotic and Parasitic Diseases, National Institute for Communicable Diseases of the National Health Laboratory Service, Johannesburg, South Africa , Johannesburg (South Africa)
  • 2 University of the Witwatersrand, Johannesburg, South Africa , Johannesburg (South Africa)
  • 3 University of Notre Dame, Notre Dame, IN, USA , Notre Dame (United States)
Type
Published Article
Journal
Malaria Journal
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Feb 24, 2020
Volume
19
Issue
1
Identifiers
DOI: 10.1186/s12936-020-03168-x
Source
Springer Nature
Keywords
License
Green

Abstract

BackgroundAccurate Anopheles species identification is key for effective malaria vector control. Identification primarily depends on morphological analysis of field samples as well as molecular species-specific identifications. During an intra-laboratory assessment (proficiency testing) of the Anopheles funestus group multiplex PCR assay, it was noted that Anopheles arabiensis can be misidentified as Anopheles leesoni, a zoophilic member of the An. funestus group. The aim of this project was, therefore, to ascertain whether other members of the Anopheles gambiae complex can also be misidentified as An. leesoni when using the standard An. funestus multiplex PCR.MethodsThe An. funestus multiplex PCR was used to amplify DNA from An. gambiae complex specimens. These included specimens from the laboratory colonies and field samples from the Democratic Republic of Congo. Amplified DNA from these specimens, using the universal (UV) and An. leesoni species-specific primers (LEES), were sequence analysed. Additionally, An. leesoni DNA was processed through the diagnostic An. gambiae multiplex PCR to determine if this species can be misidentified as a member of the An. gambiae complex.ResultsLaboratory-colonized as well as field-collected samples of An. arabiensis, An. gambiae, Anopheles merus, Anopheles quadriannulatus, Anopheles coluzzii as well as Anopheles moucheti produced an amplicon of similar size to that of An. leesoni when using an An. funestus multiplex PCR. Sequence analysis confirmed that the UV and LEES primers amplify a segment of the ITS2 region of members of the An. gambiae complex and An. moucheti. The reverse was not true, i.e. the An. gambiae multiplex PCR does not amplify DNA from An. leesoni.ConclusionThis investigation shows that An. arabiensis, An. gambiae, An. merus, An. quadriannulatus, An. coluzzii and An. moucheti can be misidentified as An. leesoni when using An. funestus multiplex PCR. This shows the importance of identifying specimens using standard morphological dichotomous keys as far as possible prior to the use of appropriate PCR-based identification methods. Should there be doubt concerning field-collected specimens molecularly identified as An. leesoni, the An. gambiae multiplex PCR and sequencing of the internal transcribed spacer 2 (ITS2) can be used to eliminate false identifications.

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