以分子生物學及病理學方法診斷禽類淋巴球增生性腫瘤疾病

禽類淋巴球增生性腫瘤性疾病，特別是淋巴瘤常發生於多種禽類品種，此些病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。這三個病毒不僅影響禽類產業且同時也造成了多種瀕臨絕種野生鳥類的滅絕，此外這三種疾病沒有典型的病理區別診斷證據可供參考，造成了診斷上極大的困難。研究上通常使用免疫組織化學染色(immunohistochemistry, IHC)、原位雜合技術（in situ hybridization, ISH）和聚合酶鏈鎖反應（polymerase chain reaction, PCR）作為診斷工具。基於上述本研究主要目的為使用PCR及IHC方式進行前述三種病毒引起之禽類淋巴瘤區別診斷。在本研究中，共收集了103個禽類淋巴瘤的組織樣本，其中包括93例石蠟包埋組織塊及10例新鮮病材；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方面顯示分別在ALV, MDV及REV檢測出55.43%, 21.36%, 和 21.36%的陽性結果。IHC方面使用病毒抗體分別在ALV, MDV及REV檢測出75.81%, 75.81%, 和 61.29% 的陽性結果。結果顯示IHC在禽類淋巴瘤之石蠟包埋組織塊是一項最有效的檢測工具，但是在發展成熟的淋巴瘤細胞卻檢測不到病毒的抗原蛋白。ISH的建立成功幫助了本研究中在IHC陰性的腫瘤細胞中偵測到REV的病毒核酸。由結果顯示，合併使用IHC及ISH技術在回溯性調查研究中為診斷禽類淋巴球增生性腫瘤性疾病最具有效性的方法。

關鍵字

禽類；淋巴球增生性腫瘤性疾病；免疫組織化學染色；聚合酶鏈鎖反應；原位雜合技術

並列摘要

Avian lymphoproliferative neoplastic diseases, especially lymphoma often occurs in a variety of avian species and have economic impact due to associated high mortality and decreased egg production rate. Most of the avian lymphomas are associated with Marek's disease virus (MDV), avian leukosis virus (ALV) or reticuloendotheliosis virus(REV). These three viruses not only influence the poultry industry but also affect a variety of endangered species of wild birds and often lack of classical pathological diagnostic feature. Therefore, immunohistochemistry (IHC), in situ hybridization (ISH), and polymerase chain reaction (PCR) have been developed as diagnostic tools. Based on the purpose aforementioned, the aim of this study is to use PCR and IHC to differentiate the three virus-induced lymphomas. In this study, a total of 103 avian lymphoma tissue samples collected including 93 cases of paraffin-embedded tissue and 10 cases of fresh samples. All samples comprised of the nervous system, cardiovascular system, gastrointestinal system, genito-urinary system and the lymphatic system. The positive results of PCR assay show 55.43%, 21.36%, and 21.36% in ALV, MDV, and REV retrospectively. The IHC using viral antibodies shows 75.81%, 75.81%, and 61.29% in ALV, MDV, and REV retrospectively. In conclusion, IHC employed in formalin-fixed paraffin-embedded tissues of avian lymphoid neoplasms is the most effective diagnostic tool among the three diagnostic tools, especially in retrospective studies; however, viral antigens are less detected among the well-developed tumor cells of ALV or REV induced lymphomas. Establishment of ISH has successfully detected the REV nucleic acid in well-developed tumor cells in this study. By the results showed that the combination of using IHC and ISH has a great potential in retrospective study for the diagnosis of the three viruses associated avian lymphoproliferative neoplasms.

並列關鍵字

Avian ； lymphoproliferative neoplastic diseases ； immunohistochemistry ； polymerase chain reaction ； in-situ hybridization

參考文獻

Aly, M. M., Smith, E. J. and Fadly, A. M. (1993). Detection of reticuloendotheliosis virus infection using the polymerase chain reaction. Avian Pathol, 22, 543-554.

Aulisio, C. G. and Shelokov, A. (1969). Prevalence of reticuloendotheliosis in chickens: immunofluorescence studies. Proc Soc Exp Biol Med, 130, 178-181.

Baba, T. W. and Humphries, E. H. (1985). Formation of a transformed follicle is necessary but not sufficient for development of an avian leukosis virus-induced lymphoma. Proc Natl Acad Sci U S A, 82, 213-216.

Baba, T. W. and Humphries, E. H. (1986). Selective integration of avian leukosis virus in different hematopoietic tissues. Virology, 155, 557-566.

Baigent, S. J. and Davison, T. F. (1999). Development and composition oflymphoid lesions in the spleens of Marek's disease virus-infected. In: Avian Pathology, Vol. 28, Taylor & Francis Ltd, pp. 287.

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以分子生物學及病理學方法診斷禽類淋巴球增生性腫瘤疾病

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