Effects of Post Ingestion and Physical Conditions on PCR

Amplification of Host Blood Meal DNA in Mosquitoes
*MA Oshaghi 1, AR Chavshin 1, H Vatandoost 1, F Yaaghoobi 1, F Mohtarami 1
M Hashemzadeh 2, N Noorjah 1, MH Modaresi 3
1Dept. of Medical Entomology, School of Public Health and Institute of Health Researches, Tehran University of
Medical Sciences, Iran
2Cellular and Molecular Research Center, Shahre Kord, Charmahal and Bakhtiary University of Medical
Sciences, Iran
3 Dept. of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Iran
(Received 3 Dec 2004; revised 3 Sep 2005; accepted 24 Sep 2005)

Abstract
Identification of host blood meal in haematophagous arthropods is an important element in their rule in transmission of
vector borne diseases. The effects of post ingestion and physical conditions that killed mosquitoes are stored on the success
of detecting blood meal DNA of Anopheles stephensi and Culex quinquefasiatus was investigated by polymerase chain reaction
(PCR) amplification at the human mitochondrial DNA cytochromeB (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 needs for meals digested longer time. Successful PCR amplification among meals digested for 36 h
dropping to a faint band. There were no differences between PCR success rate for sampled stored at +4° C or -20° C, but
less successful products were observed in samples kept at 4° C for periods longer than 30 h digestion. The results of this
study is important in malaria epidemiological studies to provide valuable information about the degree of contact between
human hosts and mosquito vectors, impact of vectors controls such bed nets and repellents, and the transmission dynamics
of human malaria and other vector-borne diseases.
Key words: Blood meal, Anopheles stephensi, Culex quinquefasiatus, CytochromeB, Malaria, Vector-borne diseases, Iran