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- A Non-Radioactive Polymerase Chain Reaction Method for Diagnosis of Machado-Joseph Disease
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題 名 | A Non-Radioactive Polymerase Chain Reaction Method for Diagnosis of Machado-Joseph Disease=非放射性聚合酶連鎖反應方法用以診斷Machado-Joseph疾病 |
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作 者 | 謝明麗; 李宣佑; 陳逸芸; 蔡嘉仁; 林玫伶; 李娟; 吳明娟; | 書刊名 | 臺灣醫學會雜誌 |
卷 期 | 98:10 1999.10[民88.10] |
頁 次 | 頁705-707 |
分類號 | 415.9 |
關鍵詞 | 非放射性聚合酶連鎖反應方法; Machado-Joseph疾病; CAG trinucleotide repeat; Machado-Joseph disease; Agarose gel electrophoresis; |
語 文 | 英文(English) |
英文摘要 | Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 (SCA3) is caused by unstable CAG trinucleotide repeat expansion in the coding region of the MJD gene. In this study we describe a non-radioactive polymerase chain reaction (PCR) method to detect the CAG repeat range of the MJD gene. This technique allows direct visualization of the PCR products on ethidium bromide-stained agarose gels within hours. In this studyl, genomic DNA samples isolated from peripheral lymphocytes, amniotic fluid cells, and chorionic villi were tested with two sets of commonly used MJD primers. PCR conditions were optimized, which resulted in clear visualization of both the primer sets on 3% agarose gels. Ten out of 25 candidate MJD patients have been identified with this method to date, with no false-positive of false-negative diagnoses. This simple, reliable, and cost-effective method can be used for patient diagnosis, pre-symptomatic diagnosis, and prenatal diagnosis. |
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