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以非生物素山葵過氧化酵素免疫組織化學染色法、聚合酶鏈鎖反應及原位雜合技術診斷禽類淋巴球增生性腫瘤疾病

Using Non-biotin HRP Immunohistochemistry, Polymerase Chain Reaction, Hybridization to Diagnose Avian Lymphoproliferative Diseases

摘要


禽類淋巴球增生性腫瘤性疾病,尤其是淋巴瘤,常發生於多種禽類品種,造成雞隻之死亡及產蛋率下降,在全球養雞產業上造成極大的經濟衝擊。大部分的禽類淋巴瘤多與馬立克病病毒(Marek's disease virus; MDV)、家禽白血病病毒(avian leukosis virus; ALV)及網狀內皮細胞增生症病毒(reticuloendotheliosis virus; REV)有關。這三種病毒不僅影響禽類產業且同時也威脅多種瀕臨絕種野生的鳥類,因為這三種疾病並無典型的病理特徵可供區別,故本研究主要目的為使用聚合酶鏈鎖反應(polymerase chain reaction; PCR)及非生物素山葵過氧化酵素免疫組織化學染色(non-biotin immunohistochemistry; non-biotin IHC)方法以建立區別診斷。本研究共收集了70個禽類淋巴瘤的組織樣本,其中包括60例石蠟包埋組織塊及10例新鮮病材;病材包括神經系統、心血管系統、消化道系統、泌尿生殖系統及淋巴系統。PCR結果顯示分別在ALV,MDV及REV檢測出45.7%,21.4%和22.9%的陽性率。IHC使用病毒抗體分別在ALV,MDV及REV檢測出71.4%,68.6%和54.3%的陽性率。結果顯示IHC在禽類淋巴瘤之石蠟包埋組織是一項較為有效的檢測工具,但是在已癌化成淋巴瘤的細胞卻檢測不到病毒的抗原蛋白。但使用原位雜合技術(in hybridization; ISH)技術卻成功協助IHC陰性的腫瘤細胞中偵測到REV的病毒核酸。由結果顯示,在回溯性調查研究中,結合使用IHC及ISH技術,大量提高禽類淋巴球增生性腫瘤性疾病病因的診斷率。

並列摘要


Avian lymphoproliferative diseases, especially lymphomas, often occur in a variety of avian species, resulting in the death of chicken, decline of egg production. Three common viral agents of Marek's disease virus (MDV), avian leukemia virus (ALV), reticuloendothelial virus (REV) are frequently isolated from the lesions. Because of lack of typical pathological features to differentiate the three diseases, the main purpose of this study is to use non-biotinylated horseradish peroxidase immunohistochemistry (non-biotin IHC), polymerase chain reaction (PCR) to establish a differential diagnosis. In this study, 70 birds with lymphomas were collected, including 60 cases with paraffin-embedded tissue samples, 10 cases with fresh materials. The sampling tissues included the nervous system, cardiovascular system, digestive system, genitourinary system, the lymphatic system. PCR results were ALV (45.7%), MDV (21.4%), REV (22.9%) detected from all 70 cases. IHC using monoclonal antibodies against the causing viruses were ALV (71.4%), MDV (68.6%), REV (54.3%) detected from all 70 cases. The results showed that IHC has a higher detection rate than PCR. However, the viral antigens using either IHC or PCR were not found in REV lesions, hybridization (ISH) has successfully detected viral nucleic acid in REV. The results show the combination of IHC, ISH employed in the retrospective study will improve definite diagnosis in avian lymphoproliferative diseases.

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