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題 名 | 雞傳染性喉頭氣管炎聚合酶連鎖反應診斷技術之建立與應用=Development and Application of Polymerase Chain Reaction on Diagnosis of Infectious Laryngotracheitis |
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作 者 | 謝快樂; 溫元文; 鄭淑文; 沈瑞鴻; 李龍湖; | 書刊名 | 臺灣畜牧獸醫學會會報 |
卷 期 | 66:2 1996.06[民85.06] |
頁 次 | 頁93-105 |
分類號 | 437.257 |
關鍵詞 | 雞傳染性喉頭氣管炎; 聚合酶連鎖反應; 雞; Infectious laryngotracheitis; Polymerase chain reaction; Chicken; |
語 文 | 中文(Chinese) |
中文摘要 | 雞傳染性喉頭氣管炎的診斷,除了靠疫學、臨床症狀、解剖及組織病理學病 變外,尚須配合一種以上的實驗室檢驗才能加以確診。為了能快速診斷此病,本實驗利用 聚合?連鎖反應來偵測傳染性喉頭氣管炎病毒核酸。雞傳染性喉頭氣管炎野外病毒株(埔里 株)的0.63kb Hind Ⅱ片斷經定序後,設定一對外側引子進行聚合?連鎖反應,其敏感度可 達lpg之病毒核酸;取部分產物以另一對內側巢式引子進行巢式聚合?連鎖反應,其敏感度 則高達l fg(約相當於6套病毒基因體所含核酸)。以6株不同的野外株及疫苗株病毒核酸 為模板進行反應,可證明其能廣泛適用於各病毒株;並以雞胚胎腎臟細胞、馬立克病毒血清 型1、2型、火雞?疹病毒、禽痘毒、產蛋下降症候群病毒以及黴漿茵之核酸為模板進行反 應,其反應具有高度特異性。此一診斷系統經動物試驗應用後證明其能簡便地自氣管檢體中 測得急性感染期與潛伏感染期之病毒核酸。故以聚合?連鎖反應偵測雞傳染性喉頭氣管炎病 毒之技術,其具有快速、高敏感性及特異性之優點,可應用於臨床雞傳染性喉頭氣管炎之診 斷。 |
英文摘要 | Diagnosis of infectious laryngotracheitis (ILT) usually relies on clinical observations, necropsy findings, and laboratory examinations. The purpose of this study was to establish a polymerase chain reaction (PCR) method for the detection of the presence of ILT virus (ILTV) in tissues. The method developed was able to detect 1 pg of viral DNA after 30 cycles of amplification. The sensitivity reached to level of 1 pg, or 6 genome equivalent of ILTV DNA, if the PCR product was re-amplified with a pair of nested primers. Sensitivity studies showed that the PCR primers were able to amplify the DNA of 4 wild types and 2 vaccine strains of ILTV. On the other hand, PCR product was not detected if the DNA isolated from chicken embryo kidney cells, Marek's disease virus serotype 1 and 2, herpesvirus of turkey, fowl poxvirus, egg drop syndrome 1976 virus, or Mycoplasma gallisepticum was used as a template. The specificity was also confirmed by using hybridization or restriction endonuclease digestion techniques. The nested PCR was also useful for the detection of both acute and latent ILTVs in the tracheal swabs. The procedure was proved to be rapid, highly specific and sensitive, and could be used for the purpose of clinical ILT diagnosis. |
本系統中英文摘要資訊取自各篇刊載內容。