Evaluation of Loop-mediated Isothermal Amplification (LAMP), Polymerase Chain Reaction (PCR), and Non-biotin Polymerized Horseradish Peroxidase (Non-biotin HRP) Immunohistochemistry for Detection of Pseudorabies Virus in Archival Paraffin Sections of Swine Nervous Tissue

利用迴路媒介恆溫增幅法、聚合酶鏈鎖反應及非生物素山葵過氧化酵素免疫組織化學染色法診斷蠟塊中豬假性狂犬病

陳幼岭(Yu-Ling Chen) ； 張清棟(Ching-Dong Chang) ； 林正忠(Cheng-Chung Lin) ； 蔡睦宗(Mu-Tsung Tsai) ； 劉振軒(Chen-Hsuan Liu)

《臺灣獸醫學雜誌》 38卷3期 (2012/09) Pp. 153-163

https://doi.org/10.7009/TVJ.201209.0153

摘要

豬假性狂犬病（Pseudorabies; PrV）為豬隻常見之神經系統性傳染病，且可能造成嚴重的神經系統疾病及導致幼豬死亡。本研究係使用保存達20年之豬神經系統疾病之蠟塊，比較迴路媒介恆溫增幅法（loop-mediated isothermal amplification; LAMP）、聚合酶鏈鎖反應（polymerase chain reaction; PCR）與非生物素山葵過氧化酵素（non-biotin horseradish peroxidase; non-biotin HRP）免疫組織化學染色法三種檢測方法之檢測結果。本實驗共收集27例豬之神經系統性疾病蠟塊（其中14例已確診為PrV感染而剩餘13例未確診）。結果顯示在14例已確診病例中共7例使用免疫組織化學染色法檢出陽性，5例使用迴路媒介恆溫增幅法檢出陽性及2例使用聚合酶鏈鎖反應檢出陽性；而在13例未確診病例中，共3例使用迴路媒介恆溫增幅法檢出陽性，2例使用聚合酶鏈鎖反應檢出陽性及1例使用免疫組織化學染色法檢出陽性。綜合結果顯示，併用迴路媒介恆溫增幅法及免疫組織化學染色法可快速並方便的檢測出蠟塊中之病原。而於田野間之假性狂犬病爆發病例檢測上，更可使用迴路媒介恆溫增幅法以大幅縮短檢測時間並提高檢出率。

關鍵字

豬假性狂犬病 ； 迴路媒介恆溫增幅法 ； 聚合酶鏈鎖反應 ； 非生物素山葵過氧化酵素免疫組織化學染色法

並列摘要

Pseudorabies is a common infectious disease in pigs and may cause severe neurological complications, resulting in fatalities in young pigs. This report is to evaluate three methods of loop-mediated isothermal amplification (LAMP), polymerase chain reaction (PCR), and non-biotin polymerized horseradish peroxidase (non-biotin HRP) immunohistochemistry to detect pseudorabies virus (PrV) in archival paraffin sections of swine nervous tissues stored up to twenty years. A total of 27 cases (14 confirmed and 13 unidentified) were used in this study. The results showed that 7/14 with non-biotin HRP, 5/14 with LAMP, and 2/14 with PCR were positive for detection of PrV in confirmed cases, and 3/13 with LAMP, 2/13 with PCR, and 1/13 with non-biotin HRP were positive for PrV in unidentified cases. The detection rate with LAMP used in either confirmed or unidentified cases is higher than the PCR assay. Based on this study, combinations of LAMP and non-biotin HRP immunohistochemistry may potentially provide a convenient and rapid detection tool for retrospective studies of viral infections. In addition, because of no use of sophisticated laboratory equipment, the LAMP assay would be advantageous for use in rapid clinical diagnosis of PrV infection in developing countries during disease outbreak.

並列關鍵字

LAMP ； non-biotin HRP immunohistochemistry ； PCR ； PrV