查詢結果分析
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頁籤選單縮合
題名 | Detection of Pseudorabies Virus Genetic Sequences by Polymerase Chain Reaction=以聚合酶連鎖反應檢測假性狂犬病毒核酸 |
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作者 | 張天傑; | 書刊名 | 臺灣畜牧獸醫學會會報 |
卷期 | 57 1991.06[民80.06] |
頁次 | 頁1-17 |
分類號 | 437.246 |
關鍵詞 | 狂犬病毒核酸; 假性; 連鎖反應; 聚合酶; 檢測; |
語文 | 英文(English) |
中文摘要 | 本試驗採用了聚合?連鎖反應以檢測假性狂犬病毒(PRV)核酸。本試驗選取了三 套引子來檢測PRV核酸並比較了各套引子的特異性。共有九株PRV及一株Cooper的牛傳染性 鼻氣管炎病毒(BHV-1)被用來測試引子的特異性。在所選用的三套引子中,BamHl-15片段引 子會使所有PRV株產生283鹼基對的產物,但亦使BHV-1產生294鹼基對的產物;若使用的 引子是來自PRV的立即早期基因時,只有UCD和SHOPE等PRV美國分離株會產生一種271鹼 基對的產物,而可被用以區分美國株及台灣株;當使用的是gX醣蛋白基因引子時,所有的 PRV病毒株皆會產生一種294鹼基對的產物,而BHV-1則無產物產生。本試驗並以gX基因引 子測試了以聚合?連鎖反應偵測PRV核酸的敏感性,當以溴化乙基啶染色時,發現可檢出的 敏感性為一個病毒感染單位(PFU)或0.1pg的PRV DNA。但若再進一步以南方轉漬雜合法檢 測時,則可提高其敏感性至0.l PFU。 |
英文摘要 | The method of polymerase chain reaction (PCR) has been adapted for the detection of genetic sequences of the psedudorabies virus (PRV). Three primer pairs of sequences were chosen and their specificities on the detection of PRV genetic sequences were compared. Nine strains of PRV and the Cooper strain of bovine herpesvirus l (BHV-1) were used for the comparison of the specificity of these primer pairs on the amplification reaction. The primer pair selected from the BamHi-L5 fragment not only produced an amplified product of 283 base pairs (bp) when reacted with the PRV strains, but also generated a product of about 194 bp on the reaction with the BHV-1 DNA. The reaction using the primer pair chosen from the immediate early gene only generated a specific PCR product of 271 bp for the UCD and SHOPE strains, but not for any of the Taiwan strains. When the primer pair sequences adapted from the gX glycoprotein gene were used, a specific DNA product of 294 bp was generated from the enzymatic amplification of all the PRV strains tested, but not of the BHV-1 or cell DNA. The sensitivity of the PCR system using gX gene as the primer pair could be as high as 1 PFU of PRV or 0.1 gp of PRV DNA by ethidium bromide staining, and 0.1 PFU by Southern blot hybridization. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。