Identification of blood meals imbibed by phlebotomine sand flies using cytochrome b PCR and reverse line blotting.
Blood meal identification is important for determining the host preferences and the vectorial capacity of hematophagous arthropods. In the past, mostly serological techniques using host-specific antibodies were used, but in recent years more sensitive and accurate polymerase chain reaction (PCR)-based molecular approaches for identifying blood meals have been developed. Here, a vertebrate-specific PCR is combined with reverse line blot analysis for identifying blood meals ingested by female phlebotomine sand fly vectors of leishmaniasis. Species-specific oligonucleotides were covalently linked to nylon membranes, and biotinylated PCR products of the mitochondrial cytochrome b gene were used as probes in a hybridization reaction revealed using colorimetric or enhanced chemiluminescent detection systems. This combination identified blood meals up to 96 hours after ingestion containing minimal amounts of DNA (>0.1 pg). The specific probes discriminated between putative host species in several study areas. The source of blood was identified in 68 of 89 wild-caught sand flies tested (76%). Mixed blood meals were identified in 15 (17%) of those. The advantages and limitations of this method are discussed.
Abbasi
Ibrahim
I
Department of Parasitology, The Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Cunio
Ruben
R
Warburg
Alon
A
D000818:Animals; D001717:Birds; D002417:Cattle; D002645:Chickens; D010856:Columbidae; D045303:Cytochromes b / Q000235:genetics*; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004197:Disease Reservoirs / Q000145:classification* / Q000469:parasitology; D005260:Female; D006801:Humans; D007043:Hyraxes; D007303:Insect Vectors / Q000469:parasitology*; D007891:Leishmania / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000635:transmission; D051379:Mice; D009693:Nucleic Acid Hybridization; D010691:Phlebotomus / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013997:Time Factors
D017931:DNA Primers; D004247:DNA; D045303:Cytochromes b