Moulay S, Zientara S, Sailleau C, Cruciere C
Centre National d'Etudes Vétérinaires et Alimentaires, Laboratoire Central de Recherches Vétérinaires, Maisons-Alfort, France.
Mol Cell Probes. 1995 Aug;9(4):233-7. doi: 10.1016/s0890-8508(95)90092-6.
In order to develop a non-radioactive dot-blot hybridization assay, for the detection of African-horse sickness virus (AHSV), genome segment 7 from 9 serotypes was amplified by RT-PCR. The resulting PCR products were denatured, immobilized on nylon membranes and then hybridized to a non-radioactive digoxigenin-labelled probe. This probe (265 bp in length) was generated by nested-PCR using genome segment 7 of AHSV, serotype 4 as a template. The dot-blot was visualized by chemiluminescence. Positives were obtained from the PCR products amplified from all 9 AHSV serotypes, but not from any other equine virus or orbivirus isolates. The sensitivity and specificity of this probe, together with the simplicity and rapidity of this technique, suggest that a non-radioactive dot blot assay may be useful as a method for the routine and rapid diagnosis of viral infections.
为了开发一种用于检测非洲马瘟病毒（AHSV）的非放射性斑点杂交检测方法，通过逆转录聚合酶链反应（RT-PCR）扩增了9个血清型的基因组片段7。将所得的PCR产物变性，固定在尼龙膜上，然后与非放射性地高辛标记的探针杂交。该探针（长度为265 bp）是以AHSV血清型4的基因组片段7为模板，通过巢式PCR产生的。通过化学发光使斑点杂交可视化。从所有9种AHSV血清型扩增的PCR产物中均获得阳性结果，但从任何其他马病毒或环状病毒分离株中均未获得阳性结果。该探针的敏感性和特异性，以及该技术的简单性和快速性表明，非放射性斑点杂交检测方法可能作为一种常规快速诊断病毒感染的方法而有用。
