Identification of mosquito avian-derived blood meals by polymerase chain reaction-heteroduplex analysis.

Abstract

Abstract. A polymerase chain reaction (PCR) heteroduplex assay (HDA) was developed to identify avian derived mosquito blood meals to the species level. The assay used primers amplifying a fragment of the cytochrome B gene from vertebrate but not invertebrate species. In Culex tarsalis fed on quail, PCR products derived from the quail cytochrome B gene were detected seven days post-engorgement. In an analysis of wild-caught mosquitoes, 85% of blood-fed mosquitoes produced detectable PCR products. Heteroduplex patterns obtained from bird-derived PCR products were found to permit the unambiguous identification of all species examined. No intraspecific variation in HDA patterns was found. The PCR-HDA was used to characterize blood meals in wild caught Cx. tarsalis. Of the 67 blood meals analyzed, 60% were derived from avian sources. Of the avian blood meals, 65% were derived from a single host, the common grackle.

Importance of Common Wall Lizards in the Transmission Dynamics of Tick-Borne Pathogens in the Northern Apennine Mountains, Italy

“…In each PCR run, distilled water was added as negative control; DNA from B. afzelii (Nancy strain) and R. conorii (Malish strain) were used as positive controls. The efficiency of the extraction protocol was verified in PCR-negative samples: for tick extracts, by a 16S rDNA PCR [6]; and for tail tissue extracts, by a cytB gene PCR [23].…”

Host Feeding Patterns of Established and Potential Mosquito Vectors of West Nile Virus in the Eastern United States

“…Two aliquots of the resulting PCR product were then separately mixed with 6 μL of two PCR product drivers derived from Northern Cardinal or Carolina Chickadee. Heteroduplex formation and separation of the heteroduplex products were carried out as previously described (Lee et al 2002). Samples were identified on the basis of a comparison of the relative mobility of the HDA bands to those of standard samples derived from avian blood DNA.…”

Community Participation and Domiciliary Occurrence of Infected Meccus longipennis in Two Mexican Villages in Jalisco State

“…The cytochrome b gene was then amplified with specific primers for the vertebrates from each DNA sample and heterodu plexes were produced with Sigmodon mascotensis amplified cytochrome b from a cloned gene in our laboratory as driver following the method previously described. 21 The heteroduplex products were formed by denaturizing at 100°C for 2.5 min of cytochrome b mixtures (V/V) of sample and driver and slow cooling of the denatured solution at room temperature. Finally, the comparison of the heteroduplex patterns was done with 10% acrylamide electrophoreses (29 acrylamide: 1 bisacrylamide) in TBE buffer.…”