腦脊髓液檢體中單純疱疹病毒、巨細胞病毒之分子診斷及化學激素成份分析

Molecular diagnosis of herpes simplex virus、cytomegalovirus and chemokine analysis in cerebrospinal fluid samples

紀有財(Yu-Tsai Chi)

指導教授 ： 黎慶 ； 陳志豪

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2005年)

https://doi.org/10.6834/CSMU.2005.00071

摘要

中樞神經系統的感染，除了細菌外病毒也是很重要的致病源。可造成中樞神經系統感染的病毒很多，除常見的腸病毒之外，單純疱疹病毒和巨細胞病毒所形成的潛伏性感染，在免疫功能不全的病人身上也可引起嚴重腦炎甚至是死亡。目前臨床檢測病毒的方法是以病毒培養和病毒特異性抗體檢測，病毒培養耗時長且常為陰性結果，而病毒抗體檢測又無法準確得知病毒復發和新興感染的真實情況，因此本實驗利用聚合酶連鎖反應來偵測病毒的存在與否，是個較快速又敏感的方法；又由許多的文獻報告中發現病毒感染中樞神經系統可引起腦脊髓液中化學激素成份之變化，於是本實驗採用酵素免疫吸附分析法檢測套組來測定含有病毒基因的腦脊髓液化學激素的含量。由實驗中得到的結果：單純疱疹病毒和巨細胞病毒陽性率分別為26.1%、27.5%，兩者都為陽性者有8.7%；化學激素的含量測定顯示這些含有病毒基因的腦脊髓液其細胞激素是增加的，其中GRO α 含量在所有陽性病例腦脊髓液中均可偵測到。另外針對陰性的腦脊髓液檢體採用序列非依賴性單一引子增幅試驗，來尋找未知病毒基因，並未明顯得到陽性結果的呈現。從實驗中得知以腦脊髓液來檢查中樞神經系統的感染有很大的診斷價值，發展一套快速偵測腦脊髓液中病毒的感染對於疾病的診治幫助很大，臨床上值得廣泛使用；而在不知名病毒的偵測方面，雖在本實驗中未獲具體的結果，這套鑑定病毒基因的系統仍值得我們更致力去建立，並將各種問題一一去克服。

關鍵字

腦脊髓液 ； 單純疱疹病毒 ； 巨細胞病毒 ； 化學激素

並列摘要

Besides bacteria, viruses are significant culprits of central nervous system infections. Many viruses cause infection of the central nervous system. In addition to the frequently-seen enterovirus, herpes simplex type I and cytomegalovirus may set off latent infection that often results in severe encephalitis or death in patients of immunodeficiency. At present clinical examination employs virus cultivation and virus distinctive antibody examine for identification of these two viruses. Virus cultivation is time-consuming and the result is often negative, while virus antibody examine is unable to accurately distinguish recurrences from new infections. Therefore this experiment employs polymerase chain reaction to detect whether viruses are present. It is a speedy sensitive method. Many literatures have documented the fact that infection of the central nervous system may result in changes in the content of chemokine of cerebrospinal fluid. So this experiment utilizes the test set of enzyme linked immunosorbent assay to determine the content of chemokine of cerebrospinal fluid that contains genes of the viruses. Experiment results: The positive rates of herpes simplex type I and cytomegalovirus in polymerase chain reaction are 26.1% and 27.5%, respectively. In 8.7% of the cases both are positive. Tests of the content of cytokine indicate the cell of virus-infected cerebrospinal fluid may increase. GRO α can be detected in cerebrospinal fluid of all positive cases. In our attempt to seek unknown viral genes in negative cerebrospinal fluid samples through sequence independent single primer amplification test, we proved the feasibility of this approach though we didn’t obtain desired positive results. From the experiment we learn that the use of cerebrospinal fluid for determination of central nervous system infections is of great diagnostic value. The development of the system for speedy identification of viral infection in cerebrospinal fluid will be of great help to diagnosis and treatment of related diseases. It should be put to extensive clinical application. This experiment did not attain tangible result in relation to identification of unknown viruses. But the system for verification of viral genes is worth our effort.

並列關鍵字

cerebrospinal fluid ； herpes simplex virus ； cytomegalovirus ； chemokine

參考文獻

王雅利. (2005).腦脊髓液中流感病毒與腸病毒分子診斷及化學激素成份分析

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