斑点杂交生物素法检测流行性出血热病毒RNA

Detection of Epidemic Hemorrhagic Fever Virus (Hantanvirus) RNA Genome with Biotin-Labeled Probe by Dot Hybridization

为寻找一种用于检测流行性出血热病毒的分子杂交方法,以生物素-7-dATP标记流行性出血热病毒(EHFV)R_(22)株M片段的cDNAR_3克隆作探针,与人源性的EHFVH-114、H-435株RNA基因组进行斑点杂交,得到阳性结果,可检出5pg的cDNA或RNA。此探针与疱疹病毒DNA不出现杂交信号。以上结果说明这种标记探针具有EHFV特异性,可以扩大应用范围,结果还表明动物源性和人源性EHFV均具有共同的保守核苷酸序列。

Dot hybridization assay was developed to detect epidemic hemorrhagic fever virus(EHFV)RNA genome using probe which biotin-7-dATP labeled R-3 cDNA clone of M fragement of EHFV R-22 strain. This probe was used to detect RNA genome of the human original EHFV H-114 and H-435 strains in cell cultures. The positive hybridization signal was obtained from cells infected EHFV H-114 and H-435 strains, 5 pg cDNA or RNA could be obtained This probe do not react with DNA of cells infected herpes simplex virus and cytomegalovirus. It was shown that this probe was specific to EHFV, and also contain common nucleotide sequence in both human and animal original EHFV.