Preservation of Field-Collected Mosquito Blood Meals.
Journal
Cold Spring Harbor protocols
ISSN: 1559-6095
Titre abrégé: Cold Spring Harb Protoc
Pays: United States
ID NLM: 101524530
Informations de publication
Date de publication:
17 Jul 2023
17 Jul 2023
Historique:
medline:
18
7
2023
pubmed:
18
7
2023
entrez:
17
7
2023
Statut:
aheadofprint
Résumé
All PCR- and DNA-based blood meal analyses require host DNA from a mosquito blood meal to be effectively preserved between the time when the specimen is collected and the extraction of DNA. As soon as a mosquito ingests blood from a host animal, digestion of host cells and cellular components within the blood meal by enzymes in the mosquito midgut begins to degrade the host DNA templates that are the targets of polymerase chain reaction (PCR) amplification. Without effective preservation, host DNA is typically undetectable by PCR 48 h after feeding, because of digestion. Preservation methods for mosquito blood meals vary in their efficacy, and the logistics of fieldwork can limit the options for preservation of blood meals and maintenance of the integrity of host DNA. This protocol describes a method of blood meal preservation that is effective, convenient, and amenable to fieldwork in remote locations where cryopreservation at -20°C or -80°C may not be feasible. It uses a Flinders Technology Associates (FTA) card, which is a chemically treated card that lyses cells and allows nucleic acids to be preserved. This method is also expected to preserve the DNA or RNA of pathogens present within the engorged mosquito abdomen, including RNA viruses.
Identifiants
pubmed: 37460150
pii: pdb.prot108290
doi: 10.1101/pdb.prot108290
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2023 Cold Spring Harbor Laboratory Press.