從心肌炎組織切片探討常見的致病DNA病毒

Identify the Common DNA Viral Pathogens from Formalin-Fixed Paraffin-Embedded Myocardium Tissues of Myocarditis

莊傑仰(Jie-Yang Jhuang) ； 孫家棟(Chia-Tung Shun)

《台灣法醫學誌》 1卷1期 (2009/06) Pp. 49-56

https://doi.org/10.6134/tjfm.2009.01.01.11

摘要

目的：由急診室中因心肌炎而求診的案例中，經由導管性心肌切片探討常見的DNA病毒爲致病因所佔的比率。
方法：收集2001年至2005年台大醫院因心肌炎入院中，心臟組織病理切片證實爲心肌炎的案例，藉由分子生物學技術去分析，找出當中源於DNA病毒感染的盛行率。
結果：共收集到24個案例，年齡層分布爲6歲到48歲。12組檢體爲實驗組，病理診斷爲心肌炎，另外12組檢體來自法醫解剖的心臟標本，死因均非爲心臟因素所造成。在12個實驗組檢體中有二個存在巨細胞病毒(human herpes virus 5, Cytomegalovirus)，比例爲16.7%(2/12)。12個對照組檢體均呈現陰性，比例爲0%(0/12)。雖然毛細管電泳結果得到的產物大小估計值和理論大小不符，但是實驗組檢體和標準病毒株的大小是相吻合。
討論：由文獻得知病理診斷爲心肌炎案例中，確實有些是源於病毒感染所造成，在台灣尤以RNA病毒較常見，至於DNA病毒在統計上並無數據。本研究從福馬林固定且經石蠟包埋的舊組織切片(Archive tissue)中研究心肌炎DNA病毒感染的情形，發現巨細胞病毒佔了16.7%。因爲舊組織切片(Archive tissue)中DNA較微量或可能變性，所以我們使用nested PCR，其靈敏度較單次PCR高。我們的檢體經nested PCR複製後，可以直接跑毛細管電泳決定產物大小和已知標準病毒的ladder比較分析，免去傳統洋菜膠電泳後還要定序才能確定病毒的問題。由此初步的結果，建議心肌炎的案例要考慮DNA病毒爲病因的可能性。

關鍵字

心肌炎 ； 疱疹病毒 ； 巨細胞病毒 ； 巢式聚合鏈連鎖反應 ； 舊組織切片

並列摘要

Purpose: This study was undertaken to evaluate the molecular analysis of endomyocardial archive tissues in identifying the possibility of common DNA viral pathogens.
Method: We collected all available clinical record and endomyocardial archive tissues for patients who had myocarditis recorded as the clinical diagnosis at the National Taiwan University Hospital from 2001 to 2005.
Result: Findings for all available patients (6 men and 6 women; median age, 26 years) with myocarditis that fulfilled the Dallas criteria were included in this study. Twelve subjects who had died naturally except heart diseases served as control group from forensic autopsy. Nested polymerase chain reaction (PCR) was used for detection of DNA viral genomes (human herpes virus 1, human herpes virus 2, Epstein-Barr virus, and human herpes virus 5) from endomyocardial biopsied tissues. DNA viral nucleic acid were found in the hearts of 2 patients (16.7%), including human herpes virus 5 (2 patient). In the control group, no viral genome was detected.
Conclusion: In patients with unexplained myocarditis, viral infection really contributes to be an important etiology. In Taiwan, RNA virus is the common etiology, but the role of DNA virus is not well-known. We identify DNA virus from the archive tissue of the patients diagnosed as myocarditis and find 16.7% cases with CMV.
We use nested polymerase chain reaction instead of single polymerase chain reaction because it may be a more sensitive and specific tool to identify viruses.
After replication, we analyze the products with capillary electrophoresis to determine the estimated size and compare them to the standard ladder. This method replaces the need to using agarose electrophoresis and sequencing, so it is more time-saving. Based on our data, we suggest taking DNA virus into consideration while facing possible etiology of myocarditis.

並列關鍵字

Myocarditis ； Herpesviridae ； Human herpes virus 5 ； Nested polymerase chain reaction ； Archive tissue

參考文獻

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