东北农业大学学报
東北農業大學學報
동북농업대학학보
JOURNAL OF NORTHEAST AGRICULTURAL UNIVERSITY
2014年
9期
100-105
,共6页
潘龙%李广兴%聂思静%蔡皓璠%李兰兰
潘龍%李廣興%聶思靜%蔡皓璠%李蘭蘭
반룡%리엄흥%섭사정%채호번%리란란
IBV%Real-time PCR%RT-PCR%噬菌体ELISA
IBV%Real-time PCR%RT-PCR%噬菌體ELISA
IBV%Real-time PCR%RT-PCR%서균체ELISA
infectious bronchitis virus%Real-time PCR%RT-PCR%phage-mediated ELISA
IBV属于冠状病毒γ群,可引起鸡呼吸道疾病,对IBV诊断方法研究广泛,建立多种方法,而噬菌体展示技术广泛用于蛋白质研究中。使用分子生物学技术和噬菌体展示技术筛选到能与IBV S1蛋白特异性结合噬菌体建立三种IBV检测方法,Real-Time PCR、RT-PCR和噬菌体ELISA。结果表明,三种方法都可有效检测出传染性支气管炎病毒(IBV),证明三种方法可靠性,对这三种方法的灵敏性进行比较,敏感性上实时Real-Time PCR法最敏感,依次为RT-PCR和噬菌体ELISA技术,检测极限分别为7.64×101、7.64×103、1.21×105 copies·μL-1,证明筛选的IBV S1蛋白特异性亲和噬菌体可用于IBV病毒诊断,具有较高特异性和敏感性,为IBV诊断提供新方法。Real-Time PCR、RT-PCR和噬菌体ELISA三种方法各具特点,可满足不同检测需要。
IBV屬于冠狀病毒γ群,可引起鷄呼吸道疾病,對IBV診斷方法研究廣汎,建立多種方法,而噬菌體展示技術廣汎用于蛋白質研究中。使用分子生物學技術和噬菌體展示技術篩選到能與IBV S1蛋白特異性結閤噬菌體建立三種IBV檢測方法,Real-Time PCR、RT-PCR和噬菌體ELISA。結果錶明,三種方法都可有效檢測齣傳染性支氣管炎病毒(IBV),證明三種方法可靠性,對這三種方法的靈敏性進行比較,敏感性上實時Real-Time PCR法最敏感,依次為RT-PCR和噬菌體ELISA技術,檢測極限分彆為7.64×101、7.64×103、1.21×105 copies·μL-1,證明篩選的IBV S1蛋白特異性親和噬菌體可用于IBV病毒診斷,具有較高特異性和敏感性,為IBV診斷提供新方法。Real-Time PCR、RT-PCR和噬菌體ELISA三種方法各具特點,可滿足不同檢測需要。
IBV속우관상병독γ군,가인기계호흡도질병,대IBV진단방법연구엄범,건립다충방법,이서균체전시기술엄범용우단백질연구중。사용분자생물학기술화서균체전시기술사선도능여IBV S1단백특이성결합서균체건립삼충IBV검측방법,Real-Time PCR、RT-PCR화서균체ELISA。결과표명,삼충방법도가유효검측출전염성지기관염병독(IBV),증명삼충방법가고성,대저삼충방법적령민성진행비교,민감성상실시Real-Time PCR법최민감,의차위RT-PCR화서균체ELISA기술,검측겁한분별위7.64×101、7.64×103、1.21×105 copies·μL-1,증명사선적IBV S1단백특이성친화서균체가용우IBV병독진단,구유교고특이성화민감성,위IBV진단제공신방법。Real-Time PCR、RT-PCR화서균체ELISA삼충방법각구특점,가만족불동검측수요。
Avian infectious bronchitis virus (IBV), which belongs to group γ coronavirus, leads to chicken respiratory disease-infectious bronchitis (IB) and causes enormous economic loss. Latest researches on IBV diagnostic methods are undertaken very extensively, and rapid and specific diagnostic methods have obvious significance on the comprehensive prevention of IB. In this paper, molecular biology and phage display technique were used to establish three kinds of IBV diagnostic methods, i.e., real time quantitative polymerase chain reaction (qRT-PCR), reverse transcription PCR (RT-PCR), and phage-mediated enzyme-linked immunosorbent assay (phage-mediated ELISA), and their characteristics were comparatively studied. The results showed that these three methods had the reliability to assay IBV specifical y, and they could detect IBV at least 7.64 × 101, 7.64 × 103, 1.21 × 105 copies·μL-1, respectively, in which qRT-PCR had the highest sensitivity. Furthermore, the high affinity phages to IBV S1 protein were employed with phage display technique to develop phage-mediated ELISA, and it had very good specificity and sensitivity for IBV diagnosis, and it is suitable to utilized veterinary clinic application.