爲建立一種(zhǒng)快速、敏感、特異的豬弓形蟲檢測方法，根據弓形蟲保守基因大月序列，設計一套特異性引物和FAM熒光素标記的MGB探中跳針；通過(guò)對(duì) P火自CR反應體系和反應條件進(jìn)行優化篩選，建立了豬弓形蟲樹吧實時(shí)熒光定量PCR檢測方法，并對(duì)此PCR檢測方法亮兒進(jìn)行了特異性、敏感性、重複性試驗放煙；利用所建立的方法對(duì)60份疑似弓形蟲感染的臨床樣(yàng)品進村村(jìn)行了檢測。結果顯示：建立的豬弓形蟲水時實時(shí)熒光定量PCR檢測方法在101~1銀些07拷貝/μL模闆範圍内有很好(訊北hǎo)的線性關系；對(duì)弓形蟲重組陽性質粒出現陽性通紅擴增信号，但對(duì)陰性對(du報算ì)照的水和其他7種(zhǒng)病原對(duì)照未擴增出特異性曲工外線；最低檢測模闆濃度爲10拷貝/μ拍學L；自60份疑似豬弓形蟲感染樣(yàng)品中檢出對站32份陽性，并且和克隆測序結果一緻。站明結果表明，本研究建立的豬弓形蟲實時(shí)熒微懂光定量PCR檢測方法可用于豬弓形蟲的快速檢測年我，從而爲豬弓形蟲病的診斷提供了間離特異、敏感、高通量的方法。
Development and A錯有pplication of a慢制 Real-time PCR M這微ethod for Detection of Toxoplas冷們ma gondii
In order to est得服ablish a rapid，sensitive a好地nd specific method for dete鐵務ction of Toxopla計和smas gondii（T. gondi），a pair of speci影聽fic primers and FAM（fluorescein）-la腦友beled MGB probes were 黑志designed based on the conserved市白 gene sequences of T. gon舊生dii. Followed by optimizing PCR re子電action system and conditi從風ons，a real-time fluorescent PCR a飛森ssay was established，and the sp工通ecificity，sensitivity and reproduc雜頻ibility test were carried out；then 6電去0 clinical samples sus舊從pected of T. gondii infection were舞工 tested by the established m通歌ethod. The results showed 市妹that the method expressed a good li紙友near relationship when the 湖玩template was within the 開國range of 101–107 copies/μL. Specificit也她y test showed that positive signal wa厭話s observed only when amplifying the秒姐 recombinant plasmids of T. go飛那ndii，rather than the water of議愛 negative contrast or othe做公r 7 pathogens；the得近 minimum concen外農tration of detection template was 10讀河 copies/μL；32 out o通時f 60 suspected samp拿我les were detected positive，and 事那the result was consistent with得遠 that of cloning化光 and sequencing. A弟綠s a conclusion，文錯the established m照花ethod in this study could b通河e used for rapid 村弟detection of T. gondii，which provide子算d a specific，sensitive and high-through街物put method for the ide器司ntification of toxopl但公asmosis.
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