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頁籤選單縮合
題 名 | 豬緊迫綜合症之基因型檢測技術探討=Study on the Genotyping of Porcine Stress Syndrome |
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作 者 | 鄭益謙; 劉名允; 劉學陶; 黎漢龍; | 書刊名 | 中華民國獸醫學會雜誌 |
卷 期 | 23:3 1997.06[民86.06] |
頁 次 | 頁274-282 |
分類號 | 437.657 |
關鍵詞 | 豬緊迫綜合症; 基因型檢測; 聚合酶鏈反應; 鹵乙烷檢測法; PSS; Genotyping; PCR; HCT; |
語 文 | 中文(Chinese) |
中文摘要 | 豬緊迫綜合症 (Porcine stress syndrome, PSS) 為調節骨骼肌細胞內鈣離子濃 度平衡之 ryanodine receptor 基因之 C1843 突變為 T1843 所導致。目前己有合併利用聚 合�t鏈反應 (Polymerase chain reaction, PCR) 和限制�t片段長度多型性 (Restriction fragment length polymorphism, RFLP) 原理發展之檢測技術可以鑑定豬之 PSS 基因型, 代替傳統使用鹵乙烷檢測法 (Halothane challenge test, HCT)。此一基因型檢測所用樣品 為抗凝之血液,再純化其白血球的去氧核醣核酸做為進行 PCR 之模版。 本文將現代操作步 驟做以下修改,如以去離子水加入血樣後加熱,利用細胞內外壓力不等張及溫度效應以製備 PCR 模版, 配合一對自行設計之引子 (Primer) 直接進行 PCR,得到 363 鹼基對長的產物 ,做為後續檢測材料; 為便利保存、運輸,同時以血液乾燥於濾紙上的採樣直接 PCR 進行 基因型檢測試驗,修改後之基因型檢測結果與目前所用者完全相同。本文並以 746 頭 8 週 齡豬採血進行基因型檢測,與 HCT 結果做比較; 共發現 23 頭豬 (3.1 % ) 判定 HCT 陽 性, 但 PSS 基因型檢測有 36 頭 T1843 純合子, 其中 19 頭為 HCT 判定陰性; 170 頭 PSS 基因型雜合子中有 6 頭 HCT 判定陽性,佔 HCT 判定陽性豬之 26 % (6/23);故基因 型檢測技術不僅比 HCT 準確,且可分辨雜合子與 PSS 基因型純合子。 |
英文摘要 | Porcine Stress Syndrome (PSS) is caused by the point mutation, C1843 to T1843, of ryanodine receptor responsible for the regulation of calcium-release channel of skeletal muscle sarcoplasmic reticulum. There has been a technique of molecular genetics, polymerase chain reaction (PCR) combining with restriction fragment length polymorphism (RFLP) developed to identify the PSS genotype of pig which was typed by halothane challenge test (HCT) before. The DNA purified from white blood cell of anticoagulant blood was utilized as the template of PCR for the genotyping technique. The typing procedures established were modified in our test to improve the typing efficiency as follows: first, blood samples were mixed with deionized water and heated to release the DNA directly as the template of PCR by osmotic and thermal effects. Secondly, the unique PCR products sized 363 b.p. defined by the two chosen primers were obtained as the material for testing the genotype of PSS. Thirdly, DNA processed from the dried blood on filter, a convenient way for sample preservation and shipping between the field and laboratory, was used to carry the PCR and genotyping directly. The PSS typing results from our modified procedures was identical with those obtained by the conventional technique. We also compared the results of genotyping from the blood with those of HCT of 746 piglets at the age of 8 weeks. Although thirty-six homozygous for T1843 by genotyping was identified, there were only 23 piglets (3.1%) determined as HCT positive. There were 19 individuals among the 36 judged as HCT negative. Six out of the 170 piglets genotyped heterozygous for PSS were determined as HCT positive. The six were the 26% (6/23) of HCT positive piglets. To identify the PSS genotype, the technique presented here is more accurate than HCT, also can clearly distinguish the heterozygous pigs from the PSS homozygotes which can't be done by HCT. |
本系統中英文摘要資訊取自各篇刊載內容。