Rapid and simple polymerase chain reaction-based diagnostic assays for GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japanese domestic cats
Rapid and simple polymerase chain reaction-based diagnostic assays for GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japanese domestic cats
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DOI：
10.1177/104063871102300224
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发表时间：
2011-03-01
影响因子：
1.5
通讯作者：
Yamato, Osamu
中科院分区：
文献类型：
--
作者：
Rahman, Mohammad M.;Shoubudani, Tomoaki;Yamato, Osamu
建立了基于聚合酶链反应（PCR）结合微芯片电泳的日本家猫GM2神经节脂质病变异0 （sandhoff样病）的诊断和基因分型方法。对日本南部的日本家猫种群（共1015只猫）进行了初步的基因分型调查。采用血染的Flinders Technology Associates滤纸（FTA卡）为模板，进行PCR引物诱导限制性内切分析（PIRA）和突变分离(MS)-PCR三种检测。用琼脂糖凝胶和芯片电泳对PCR产物进行分析。所有的分析都足以确定这种疾病的基因型，但MS-PCR提供了最快速和最简单的测试，因为它不需要PCR-PIRA所需的限制性内切酶步骤。使用微芯片电泳结合FTA卡片取样可以缩短基因分型所需的时间，并简化程序。在目前的研究中，基因分型调查没有发现任何猫拥有突变等位基因，这表明该等位基因的流行率很低(
Polymerase chain reaction (PCR)-based assays combined with microchip electrophoresis were developed and evaluated for diagnosis and genotyping of GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japanese domestic cats. A preliminary genotyping survey was carried out in the population of Japanese domestic cats (1,015 cats in total) in southern Japan. Three kinds of assays including PCR primer-induced restriction analysis (PIRA) and mutagenically separated (MS)-PCR were carried out using blood-stained Flinders Technology Associates filter papers (FTA cards) as templates. The PCR products were analyzed by both agarose gel and microchip electrophoreses. All assays were sufficient to determine the genotypes of this disease, but MS-PCR offered the most rapid and simplest test, as it does not need the restriction enzyme step required in PCR-PIRA. The use of microchip electrophoresis in combination with FTA cards for sampling could shorten the time required for genotyping and simplify the procedure as well. The genotyping survey in the current study did not find any cats that possessed the mutant allele, suggesting that the prevalence of this allele is low (