2013-11-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/692594摘要：家禽流行性感冒可感染人與動物。2012年3月台灣低病原性的H5N2病毒已證實突變為高病原性，2013年5月更發生全球第一例人類感染H6N1，顯示家禽流行性感冒病毒在台灣本土持續不斷突變並潛在跨物種感染的實力。2013年3月中國大陸發生人類感染H7N9死亡案例，4月則發生H7N9境外移入台灣案例，引發民眾的惶恐。現行市面上血清學的快速診斷試劑，雖可在現場第一線使用，然而敏感度過低，而高敏感度的核酸診斷法則需要專業的實驗室與技術人員，難以在現場推廣使用，故有進一步開發高效率高專一性高敏感度檢驗方法的必要，以支應瞬息萬變的病毒再重組以及現場大量病毒篩檢檢的防疫工作量。環狀等溫增幅反應，不需要PCR增幅器，可直接用肉眼判讀檢測結果；而生物晶片藉由與探針專一性的結合，則可迅速得到大量的基因分子訊息。兩者兼具有高專一性高敏感度的特質。本研究希望結合環狀等溫增幅反應與生物晶片的技術，快速檢測台灣主要的家禽流行性感冒病毒亞型H6N1, H5N2，並與H7N9甚或與人類的H6N1作鑑別診斷。全程反應不需要PCR增幅器，也不需要電泳膠與成像照膠系統，在禽場第一線即可操作，3小時內即可判讀結果。簡易快速且具備加乘提高敏感度與專一度的性質，解決目前現場血清學檢測敏感度不佳以及專業實驗室高設備與人力成本的問題，並可應付大量檢體篩檢的需求。本研究預計可架構一新興產業技術應用於台灣重要家禽流行性感冒病毒亞型之偵測與鑑別診斷。研究的成果將移轉業界，並計畫進一步商品化，包裝為生物檢驗試劑套組，初期將推往政府各地方的防檢單位以及養雞業者。預期可增加國內生技產業界的技術深度，並有助於提升國內禽畜產業界的產值，一舉增進台灣此雙相產業的競爭能力。<br> Abstract: Avian influenza virus (AIV) can infect both human and animal. The low pathogenicity avian influenza virus (LPAIV) H5N2 in Taiwan has been proven to be highly pathogenic since March of 2012. Moreover, the first global case of human H6N1 was just reported in Taiwan in May of 2013. Both of these manifest that avian influenza virus keeps mutating continuously and even the LPAIV strain has gained the ability to spread to human. There have been a number of deadly cases caused by H7N9 in China since March of 2013. The importation of H7N9 was also reported in April of 2013, which caused a panic in Taiwan. Serologic detection of the influenza virus is available in the market, but the sensitivity is low. Virus RNA detection is highly sensitive, but it needs professional lab and skilled technician and cannot be employed in the field instantly. At the moment that virus is constantly undergoing recombination, a method with higher sensitivity, higher specificity and easier approach is therefore needed to develop to bear the huge workload of virus screening. Loop-mediated isothermal amplification (LAMP) can be worked without PCR machine, and the result can be read with naked eyes. Biochip can attain lots of genetic messages at the same time by the hybridization of probes and virus neucleotides. Both of these detection instruments own high specificity and sensitivity. In this study, we would like to develop a novel detection system which combines the techniques of LAMP and biochip to detect the prevailing strains of H6N1 and H5N2 in Taiwan and differentiate them from H7N9 and human H6N1. The developed system does not need PCR machine, electrophoresis and gel imaging system, and the results can be read in the field within 3 hours. The multiplied sensitivity and specificity, easy implementation and rapid procedure could solve the problems with the methods now are currently used. The results of this study will be handed over to manufacturer, and packed into a commercial kit which could be promoted to animal disease control centers and poultry farmers. Furthermore, it could bring profits in both biotechnological and fowl industries, and elevate the industrial competitiveness in Taiwan.家禽流行性感冒環狀等溫增幅反應生物晶片Avian influenzaLoop-mediated isothermal amplificationbiochip