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嗜血昆虫的血餐鉴定是流行病学媒介评价的一个重要内容。以往用血清学方法,近年来DNA杂交已广泛用于生物体内异种DNA的鉴定,DNA探针可避免免疫学分析中固有的交叉反应,且能检测室温保存1年以上蚊胃血标本。但同位素~(32)P标记探针的半衰期短,易污染环境,因此,作者用生物素化人DNA探针斑点杂交法来鉴定蚊胃血,以适用于常规野外调查和现场应用。DNA探针为一个1.7kb的HindIII/EcoRI酶切片段,构建自人β-球蛋白基因3’端的Kpn-I重复序列(4.3kb),用Bjo-11-UTP以切口平移法标记该片段。生物素化探针的敏感性和特异性;人和猴、犬、牛、猪、小鼠、山羊6种动物的抗凝血样经裂解缓冲
Blood meal identification of bloodthirsty insects is an important part of the evaluation of epidemiological media. In the past, with serological methods, DNA hybridization has been widely used for the identification of heterologous DNA in vivo in recent years. DNA probes can avoid the cross-reaction inherent in immunological analysis and can detect mosquito-stomach blood samples stored at room temperature for more than one year. However, the isotope ~ (32) P-labeled probe has a short half-life and is easy to pollute the environment. Therefore, the authors identified the mosquito and stomach blood with a biotinylated human DNA probe dot blot hybridization for routine field investigation and field application. The DNA probe is a 1.7 kb HindIII / EcoRI fragment, constructed from the Kpn-I repeats (4.3 kb) at the 3 ’end of the human β-globin gene and nicked by Bjo-11-UTP. The sensitivity and specificity of the biotinylated probe; anticoagulated blood samples from humans and from 6 species of animals such as monkeys, dogs, cattle, pigs, mice and goats