动物医学进展
動物醫學進展
동물의학진전
PROGRESS IN VETERINARY MEDICINE
2015年
2期
11-15
,共5页
吴嫒琼%谢芝勋%胡庭俊%谢丽基%罗思思%邓显文%谢志勤%黄莉%黄娇玲%曾婷婷
吳嬡瓊%謝芝勛%鬍庭俊%謝麗基%囉思思%鄧顯文%謝誌勤%黃莉%黃嬌玲%曾婷婷
오애경%사지훈%호정준%사려기%라사사%산현문%사지근%황리%황교령%증정정
H4亚型%AIV%套式RT-PCR%检测方法
H4亞型%AIV%套式RT-PCR%檢測方法
H4아형%AIV%투식RT-PCR%검측방법
H4 subtype of AIV%nested RT-PCR%detection
本研究旨在建立一种快速、准确检测 H4亚型禽流感病毒（AIV ）的方法，针对 H4亚型AIV HA基因保守序列，设计并筛选出2对特异性引物，通过优化反应条件，建立了H4亚型AIV套式RT‐PCR检测方法。特异性试验结果显示，该法可特异性检测出 H4亚型AIV ，对其他常见禽病病原体无交叉；敏感性试验结果显示，该法对 H4亚型AIV检测下限为36．0 fg／μL ；最后用该法对106份临床样品进行检测，临床样品检测结果与病毒分离结果一致。本研究建立的套式RT‐PCR方法为 H4亚型AIV的早期诊断及有效防治提供了快速、特异和敏感的检测方法。
本研究旨在建立一種快速、準確檢測 H4亞型禽流感病毒（AIV ）的方法，針對 H4亞型AIV HA基因保守序列，設計併篩選齣2對特異性引物，通過優化反應條件，建立瞭H4亞型AIV套式RT‐PCR檢測方法。特異性試驗結果顯示，該法可特異性檢測齣 H4亞型AIV ，對其他常見禽病病原體無交扠；敏感性試驗結果顯示，該法對 H4亞型AIV檢測下限為36．0 fg／μL ；最後用該法對106份臨床樣品進行檢測，臨床樣品檢測結果與病毒分離結果一緻。本研究建立的套式RT‐PCR方法為 H4亞型AIV的早期診斷及有效防治提供瞭快速、特異和敏感的檢測方法。
본연구지재건립일충쾌속、준학검측 H4아형금류감병독（AIV ）적방법，침대 H4아형AIV HA기인보수서렬，설계병사선출2대특이성인물，통과우화반응조건，건립료H4아형AIV투식RT‐PCR검측방법。특이성시험결과현시，해법가특이성검측출 H4아형AIV ，대기타상견금병병원체무교차；민감성시험결과현시，해법대 H4아형AIV검측하한위36．0 fg／μL ；최후용해법대106빈림상양품진행검측，림상양품검측결과여병독분리결과일치。본연구건립적투식RT‐PCR방법위 H4아형AIV적조기진단급유효방치제공료쾌속、특이화민감적검측방법。
In order to develop a simple and quick method to detect H 4 subtype of avian influenza virus (AIV) ,two pairs of primers were designed based on the HA gene sequences of AIV H4 subtype .A nested RT‐polymerase chain reaction (nRT‐PCR) method was established for detection of H4 subtype of AIV .To further examine reliability ,specificity and sensitivity of the assay were evaluated ,and 106 clinical samples were tested by nested RT‐PCR .The results show that this nested RT‐PCR was specific to the H4 subtype of AIV ,without amplification of genomic materials other avian pathogens ;The detection limit of nested RT‐PCR was 36 .0 fg/μL ;The results of nRT‐PCR for detection of clinical samples were coincident with that of the viral isolation completely .Our results demonstrated that the optimized nested RT‐PCR assay was quick ,specific and sensitive for detection of H4 subtype of AIV .
