Identification of Anopheles sinensis blood meals with PCR
Author of the article:ZHU Chang-Qiang1** HAN Kun2 AI Le-Le1 QIANG Shi-Gui1 HAN Zhao-Jiu1JIANG Zhi-Kuan1 TAN Wei
Author's Workplace：1.Central of Disease Prevention and Control of Nanjing Military Area, Nanjing 21002, China; 2.Ningxia Centers for Disease Prevention Control, Yinchuan 750004, China
Key Words：Anopheles sinensis, mosquitoes blood meals, PCR, blood-sucking habit
Abstract：[Objectives] To develop a PCR method for identifying the source of Anopheles sinensis blood meals and determining the host preferences of A.sinensis. [Methods] Species-specific primers were designed based on the mitochondrial cytochrome b sequences of common mosquito hosts (human, cow, pig, goat, mouse and chicken) to identify the animal blood ingested by wild-caught A. sinensis. Meanwhile, DNA was extracted from the blood of common mosquito hosts, and from A.sinensis that had not ingested a blood meal, to verify the specificity of the method. DNA extracted from A. sinensis mosquitoes 6, 12, 24, 36, 4 h and 60 h after ingesting a blood meal was analyzed to determine the sensitivity of the method. [Results] Specific PCR products (689, 453, 225, 485, 266 and 468 bp) were amplified from the DNA extracted from different animal hosts. Host DNA was detectable in frozen mosquitoes for as long as 36 h after ingestion but could not be amplified 48 h and 60 h after ingestion. PCR analysis of the DNA in wild-caught specimens indicates that A. sinensis fed mainly on the blood of humans (50%), followed by that of pigs (30%) and cows (20%). A mixed blood meal comprised of both human and pig blood was detected in a single mosquito. [Conclusion] The PCR assay proved an accurate and reliable method of identifying the blood meals of wild-caught A. sinensis. A. sinensis feeds on both human and animal blood.