Identification of Blood Meals Imbibed by Phlebotomine Sand Flies Using Cytochrome b PCR and Reverse Line Blotting

被引:67
作者
Abbasi, Ibrahim [1 ]
Cunio, Ruben [1 ]
Warburg, Alon [1 ]
机构
[1] Hebrew Univ Jerusalem, Hadassah Med Sch, Dept Parasitol, Kuvin Ctr Study Trop & Infect Dis, IL-91120 Jerusalem, Israel
关键词
Leishmania; Sand fly (flies); Zoonosis; IMMUNOSORBENT-ASSAY ELISA; CUTANEOUS LEISHMANIASIS; BETA-THALASSEMIA; NORTHERN ISRAEL; RESERVOIR HOSTS; DIPTERA; HYBRIDIZATION; SANDFLIES; DISEASES; VECTORS;
D O I
10.1089/vbz.2008.0064
中图分类号
R1 [预防医学、卫生学];
学科分类号
1004 ; 120402 ;
摘要
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.
引用
收藏
页码:79 / 86
页数:8
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