Effects of post-ingestion and physical conditions on PCR amplification of host blood meal DNA in mosquitoes.
The effects of post-ingestion and physical conditions under which killed mosquitoes are stored on the success of detecting blood meal DNA of Anopheles stephensi and Culex quinquefasiatus were investigated by polymerase chain reaction (PCR) amplification at the human mitochondrial DNA cytochrome b (cytB) gene. Host DNA extracted from the blood meal up to 33 h post-ingestion in both species acts as an efficient template for PCR amplification. However, more DNA concentration is needed for meals digested for a longer time, and successful PCR amplification from meals digested for 36 h,dropped to a faint band. There were no differences between PCR success rate for samples stored at +4 or -20 degrees C, but less successful products were observed in samples kept at 4 degrees C for the periods longer than 30 h digestion. The results of this study are important for conducting malaria epidemiological studies that provide information about the degree of contact between human hosts and mosquito vectors, impact of vector controls such as bed nets and repellents, and the transmission dynamics of human malaria and other vector-borne diseases.
Oshaghi
Mohammad Ali
MA
Department of Medical Entomology, School of Public Health and Institute of Health Researches, Tehran University of Medical Sciences, Iran. oshaghima@gmail.com
Chavshin
Ali Reza
AR
Vatandoost
Hassan
H
Yaaghoobi
Fatemeh
F
Mohtarami
Fatemeh
F
Noorjah
Nahid
N
D000818:Animals; D000852:Anopheles / Q000502:physiology*; D003465:Culex / Q000502:physiology*; D045303:Cytochromes b / Q000235:genetics; D004272:DNA, Mitochondrial / Q000097:blood*; D004063:Digestion / Q000502:physiology; D005247:Feeding Behavior; D005260:Female; D006801:Humans; D007303:Insect Vectors / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors
D004272:DNA, Mitochondrial; D045303:Cytochromes b