摘要
应用光敏生物素标记我国流行性出血热病毒R22株M片段R3克隆cDNA,制备出生物素化的cNNA探针。用此探针与流行性出血热病毒R22株和陈株进行斑点杂交,结果R3克隆与R22株和陈株均出现阳性杂交信号,而与单纯疱疹病毒、柯萨奇B组病毒和正常对照细胞组的杂交结果均为阴性。光敏生物素标记探针可检出10pg的cDNA,并用此方法检测TEHF病毒在感染乳鼠体内的分布情况。
Photobiotin was used to prepare biotinylated nonradioactive nucleic acid probe for the detection of the Hantaviruses RNA. The R3 cDXA clone of genome iragment of R22 virus isolated from Rattus norvegicus in China was biotinylated and used as a probe to hybridize with R22 and Chen strains of hantavirus. After hybridization,biotin-labelled cDNA probe bound to viral RNA on the nitrocellulose paper was detected with an avidin-alkaline phosphatase conjugate. Haatayirus RXA was readily detected, but herpes simplex virus DNA, enteroviruses RNA and normal cell nucleic acid gave colourless spots. The sensitivity of photobiotin-labelled cDNA probe was similar to that of biotin-labelled probe, but lower than that 01 α-32P labelled probe. Hantaviruses RNA was detected in some organs of the infected mice with dot blot hybridization using photobiotin labelled cDNA probe.
出处
《病毒学报》
CAS
CSCD
北大核心
1990年第4期358-362,共5页
Chinese Journal of Virology
关键词
流行性出血热
病毒
光敏生物素
Photobiotin cDNA probe Dot blot hybridizationHantavirus
