本文選題：長鏈PCR + 病毒滅活��； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2006年碩士論文

【摘要】：目的：HBV、HCV和HIV等經(jīng)血傳播的病毒對人類危害極大，我國僅HBV感染者就超過1.2億。由于病毒感染窗口期的存在、新的病毒株不斷出現(xiàn)以及檢測技術(shù)還不完善等原因，通過輸血或血液制品傳播病毒性疾病的現(xiàn)象時有發(fā)生。為提高血液的安全性，，有必要對其進行病毒滅活處理。在血液病毒滅活研究工作中，常用細胞感染法評價病毒的滅活效果。該法直接、客觀，能夠真實反映病毒失活的程度。但由于HBV和HCV等一些病毒目前還無法在體外培養(yǎng)復(fù)制，病毒滅活效果難以用細胞感染法進行直觀評價，真實反映這些病毒的滅活效果仍是一個難題。本研究目的是探討長鏈核酸擴增技術(shù)用于評價病毒滅活效果的可行性，探索病毒滅活評價的新方法。 方法：以pTAL-Seap質(zhì)粒作為模型，通過特異性酶切(BSPH-Ⅰ)及非特異性的~(60)Co-γ射線輻照(30kGy)損傷后行不同長度核酸片段(0.3kb、0.8kb、1.7kb、2.7kb、4.8kb)的PCR擴增，探討長鏈核酸擴增技術(shù)評價DNA損傷的可行性。以具有體外致細胞病變效應(yīng)(CPE)的偽狂犬病毒(Pseudorabie virus，PRV)作為試驗病毒。PRV大量復(fù)制后經(jīng)~(60)Co-γ射線輻照、低pH值(4.0±0.1)處理、巴氏法消毒(60±1.0℃)、有機溶劑／洗滌劑法(S／D法，0.3％TNBP+1％TWEEN-80)處理以及亞甲藍／光化學(xué)法(MB／L，1.0uM亞甲藍，40000Lux光照)滅活，提取
[Abstract]:Objective\ Because of the existence of virus infection window period, the emergence of new virus strains and the imperfect detection techniques, the phenomenon of transmission of viral diseases through blood transfusion or blood products occurs from time to time. In order to improve the safety of blood, it is necessary to inactivate the virus. In the research of blood virus inactivation, the inactivation effect of virus was evaluated by cell infection method. The method is direct, objective and can reflect the degree of virus inactivation. However, some viruses such as HBV and HCV can not be cultured and duplicated in vitro, so it is difficult to evaluate the inactivation effect by cell infection. It is still a difficult problem to truly reflect the inactivation effect of these viruses. The purpose of this study was to explore the feasibility of long chain nucleic acid amplification (LSTNA) in evaluating the inactivation effect of viruses and to explore a new method for evaluating virus inactivation. Methods: pTAL-Seap plasmid was used as a model. The DNA damage was detected by PCR amplification of different length nucleic acid fragments (0.3 kb ~ (0.8) kb ~ (2. 7) kb ~ (4) kb) after specific enzyme digestion (BSPH- 鈪

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