Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, China.
J Clin Microbiol. 2012 Mar;50(3):619-25. doi: 10.1128/JCM.00848-11. Epub 2011 Dec 21.
An oligonucleotide microarray, LyssaChip, has been developed and verified as a highly specific diagnostic tool for differentiation of the 7 major lyssavirus species. As with conventional typing microarray methods, the LyssaChip relies on sequence differences in the 371-nucleotide region coding for the nucleoprotein. This region was amplified using nested reverse transcription-PCR primers that bind to the 7 major lyssaviruses. The LyssaChip includes 57 pairs of species typing and corresponding control oligonucleotide probes (oligoprobes) immobilized on glass slides, and it can analyze 12 samples on a single slide within 8 h. Analysis of 111 clinical brain specimens (65 from animals with suspected rabies submitted to the laboratory and 46 of butchered dog brain tissues collected from restaurants) showed that the chip method was 100% sensitive and highly consistent with the "gold standard," a fluorescent antibody test (FAT). The chip method could detect rabies virus in highly decayed brain tissues, whereas the FAT did not, and therefore the chip test may be more applicable to highly decayed brain tissues than the FAT. LyssaChip may provide a convenient and inexpensive alternative for diagnosis and differentiation of rabies and rabies-related diseases.
一种寡核苷酸微阵列（LyssaChip）已经被开发出来，并被验证为一种高度特异性的诊断工具，可用于区分 7 种主要的狂犬病病毒。与传统的分型微阵列方法一样，LyssaChip 依赖于编码核蛋白的 371 个核苷酸区域中的序列差异。该区域使用嵌套的反转录-PCR 引物进行扩增，这些引物结合到 7 种主要的狂犬病病毒上。LyssaChip 包括 57 对种型分型和相应的对照寡核苷酸探针（寡探针）固定在玻璃载玻片上，它可以在 8 小时内分析单个载玻片上的 12 个样本。对 111 份临床脑组织标本（65 份来自疑似狂犬病的动物，46 份来自餐馆收集的屠宰狗脑组织）的分析表明，芯片方法的敏感性为 100%，与“金标准”（荧光抗体试验）高度一致。芯片方法可以检测到高度腐烂的脑组织中的狂犬病病毒，而荧光抗体试验则不能，因此芯片试验可能比荧光抗体试验更适用于高度腐烂的脑组织。LyssaChip 可能为狂犬病和狂犬病相关疾病的诊断和鉴别提供一种方便、廉价的替代方法。
