第四十九卷(2020)
44.即時聚合酶鏈鎖反應檢測努比亞山羊黏多醣症之基因型分析
朱家德(1) 林德育(1) 賴永裕(1) 陳若菁(1) 吳明哲(1) 張秀鑾(2)
(1)行政院農業委員會畜產試驗所 (2)國立屏東科技大學
山羊黏多醣症（Mucopolysaccharidosis, MPS）IIID型，是一種遺傳缺陷所導致的代謝性疾病。黏多醣學名為醣胺多醣glycosaminoglycans（GAGs），是體內含量最豐富的多醣體；因GAGs的溶液態具有高黏度，故稱為黏多醣mucopolysaccharide。目前僅有單一品種努比亞（Nubian）山羊的GNS（N-acetylglucosamine-6-sulphatase,又稱 G6S）基因在cDNA第322個核苷酸有一突變點（C→T）。本試驗山羊以5場民間種羊場業者提供90頭努比亞種羊血液為樣本，運用螢光引子（FAM/HEX-labelled Primer）標記之即時聚合酶鏈鎖反應進行G6S基因型鑑別，分析顯示G6S正常型、雜合型及有病型分別為72.2%（65/90）、27.8%（25/90）及0%（0/90），並進行突變點拆離式聚合酶鏈鎖反應（MS-PCR）檢測與基因片段定序（DNA Sequencing）比對作業，發現3者判定之G6S基因型結果相符合。綜合上述，以即時聚合酶鏈鎖反應檢測技術做為判斷努比亞山羊黏多醣症之正常型及有病型個體，可為另一種迅速判定山羊GNS黏多醣症基因型之檢測模式。
關鍵語：山羊、黏多醣症、即時聚合酶鏈鎖反應
Genotypic Analysis of Caprine Mucopolysaccharidosis Type IIID by Real-time PCR Platform
C. T. Chu(1), D. Y. Lin(1), Y. Y. Lai(1), J. C. Chen(1), M. C. Wu(1) and H. L. Chang(2)
(1)Taiwan Livestock Research Institute (2)National Pingtung University of Science and Technology
The mucopolysaccharidoses (MPSs) are a group of lysosomal storage disorders caused by deficiency of enzymes catalyzing the stepwise degradation of glycosaminoglycans (GAGs). A model of a Nubian goat with biochemical abnormalities and an enzyme deficiency resembling was found in the Sanfilippo syndrome type D, mucopolysaccharidosis IIID (MPS IIID). In Nubian goat MPS IIID, the G6S deficiency is associated with a single mutation, changing a C to T in the 322 nucleotide of the G6S cDNA sequence. Blood and DNA were obtained from five different farms (n=90) and were tested the G6S deficiency by Real-time PCR platform. The results showed the frequencies of AA, AB, and BB genotypes were 72.2% (65/90), 27.8% (25/90), and 0% (0/90), respectively. The polymorphism (C to T) detected by Mutagenically Separated PCR and DNA sequencing had the same genotypes by Real-time PCR. The genotype of caprine mucopolysaccharidosis type IIID by Real-time PCR could be one of the genotyping methods to effectively detect the G6S deficiency in Nubian goat.
Key Words: Nubian goat, G6S, Real-time PCR
  
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