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A Simple Method for Storing Mosquito Bloodmeals for Human DNA Profiling

  • David O. OdongoEmail author
  • Lucy W. Irungu
Research Article

Abstract

A simple method for storing mosquito bloodmeal samples, which permits extraction and detection of human DNA after polymerase chain reaction (PCR) amplification of target DNA sequences, was tested. Abdomens of bloodfed field-collected Anopheles gambiae s.l. and An.fiinestus mosquitoes (Diptera: Culicidae) were directly expressed onto filter paper, air-dried and stored at room temperature. DNA was extracted and amplified at human hypervariable loci TC11, VWA and D1S80. The amplified products were separated using Polyacrylamide gel electrophoresis, visualised by silver staining, and results compared with those from mosquitoes that had been preserved in liquid nitrogen. DNA from blooded abdomens stored on dried filter papers could be amplified with greater than 95 % success for any locus, storage temperature, mosquito species or storage duration. Collection and drying of mosquito bloodmeals directly onto filter paper appears to be a more convenient method for sample transportation and storage than the conventional method involving cryopreservation.

Key Words

hypervariable loci DNA profiling mosquitoes Anopheles spp. bloodmeals PCR 

Résumé

Nous avons testé une méthode simple de conservation d’échantillons de repas de sang qui permette l’extraction et la détection d’ADN humain après amplification par PCR de séquences cibles. Les abdomens de moustiques post-prandiaux collectés sur le terrain (Anopheles gambiae et Anopheles fiinestus) sont exprimés directement sur un papier filtre, puis sont séchés à l’air et conservés à température ambiante. L’ADN est alors extrait et amplifiés aux loci hypervariables TOI, VWA, et D1S80. Les produits amplifiés sont séparés par électrophorèse sur gel de Polyacrylamide et visualisés par coloration aux sels d’argent. Les résultats ont été comparés à ceux obtenus à partir de moustiques cryoconservés en azote liquide. Ils montrent que l’ADN d’abdomens contenant du sang conservés sur papier filtre eut être amplifié avec un taux de réussite excédant 95 % quelque soient le locus, la température de conservation, l’espèce de moustique considérée ou la durée de conservation. Le prélèvement de repas de sang de moustique directement sur papier filtre apparaît donc comme une méthode pratique pour le transport et la conservation des échantillons, sans avoir recours à la cryoconservation ou toute autre manipulation sur le terrain.

Mots Clés

loci hypervariables profil d’ADN moustiques Anopheles spp. repas de sang PCR 

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Copyright information

© ICIPE 2002

Authors and Affiliations

  1. 1.Department of ZoologyUniversity of NairobiNairobiKenya

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