摘要
为评价异羟基洋地黄毒甙配基(Digoxigenin)探针(下简称Dig探针)与生物素(Biotin)探针的敏感性,本文采用斑点杂交及组织切片的原位杂交两种方法检测流行性出血热病毒(EHFV)核酸。将已知含量的EHFV cDNA点在硝酸纤维膜上,分别用Dig探针及Biotin探针与其杂交。用此方法Dig探针能测出0.1Pg EHFV cDNA,而Biotin探针只能测出1.0pg的FHFVcDNA。两种探针与含有EHFV长爪沙鼠脑的石蜡切片作原位杂交,结果Dig探针杂交阳性信号明显比Biotin探针强。表明Dig探针比Biotin探针敏感。
In order to evaluate the sensitivity of hybridi- zation using digoxigenin-and biotin-labelled probes, the EHFV nucleic acid was examined by dotblot and in situ hybridization with these two kinds of probe. A series of diluted EHFV cDNA were spo- tted onto the nitrocellulose filters and alloved to hybridize with digoxigenin-and biotin-labelled probes, resqectively. By this method, probe labelled with digoxigenin detected 0.1 pg of EHFV cDNA, probe biotinylated could only detect 1.0 pg of EHFV cDNA. Brain tissue paraffin-embedded sections of sukling meriones ungniculatus infected by EHFV were hybridized both with digoxigenin- and biotin -labelled probes. The positive signals in nerve cells were stronger with digoxigenin-labelled probe than with biotin-labelled probe. These demonstrate that hybridization with digoxigenin-labelled probe is more sensitive than the method with biotinylated probe.
