动物医学进展
動物醫學進展
동물의학진전
PROGRESS IN VETERINARY MEDICINE
2015年
1期
11-14,15
,共5页
徐倩%谢芝勋%谢丽基%邓显文%谢志勤%罗思思%黄莉%黄娇玲%曾婷婷
徐倩%謝芝勛%謝麗基%鄧顯文%謝誌勤%囉思思%黃莉%黃嬌玲%曾婷婷
서천%사지훈%사려기%산현문%사지근%라사사%황리%황교령%증정정
禽流感病毒%H9亚型%N2亚型%双重逆转录聚合酶链式反应
禽流感病毒%H9亞型%N2亞型%雙重逆轉錄聚閤酶鏈式反應
금류감병독%H9아형%N2아형%쌍중역전록취합매련식반응
Avian influenza virus%H9 subtype%N2 subtype%duplex RT-PCR
为建立简便快速检测H9及N2亚型禽流感病毒（AIV ）的方法，根据AIV H9和N2基因序列，分别设计了2对针对H9亚型AIV的HA基因和N2亚型AIV的NA基因的引物，建立了 H9亚型和N2亚型AIV双重RT‐PCR检测方法。对H9N2亚型AIV的RNA模板进行RT‐PCR扩增，可得到545 bp H9基因条带和341 bp N2基因的特异性条带；对非H9亚型的N2亚型AIV进行扩增，则仅出现1个特异性扩增条带，即341 bp N2基因条带；对非H9或N2亚型AIV和其他禽呼吸道病原体进行PCR扩增，结果均为阴性；结果表明该双重RT‐PCR最低能检出100 fg H9N2亚型AIV的cDNA模板。
為建立簡便快速檢測H9及N2亞型禽流感病毒（AIV ）的方法，根據AIV H9和N2基因序列，分彆設計瞭2對針對H9亞型AIV的HA基因和N2亞型AIV的NA基因的引物，建立瞭 H9亞型和N2亞型AIV雙重RT‐PCR檢測方法。對H9N2亞型AIV的RNA模闆進行RT‐PCR擴增，可得到545 bp H9基因條帶和341 bp N2基因的特異性條帶；對非H9亞型的N2亞型AIV進行擴增，則僅齣現1箇特異性擴增條帶，即341 bp N2基因條帶；對非H9或N2亞型AIV和其他禽呼吸道病原體進行PCR擴增，結果均為陰性；結果錶明該雙重RT‐PCR最低能檢齣100 fg H9N2亞型AIV的cDNA模闆。
위건립간편쾌속검측H9급N2아형금류감병독（AIV ）적방법，근거AIV H9화N2기인서렬，분별설계료2대침대H9아형AIV적HA기인화N2아형AIV적NA기인적인물，건립료 H9아형화N2아형AIV쌍중RT‐PCR검측방법。대H9N2아형AIV적RNA모판진행RT‐PCR확증，가득도545 bp H9기인조대화341 bp N2기인적특이성조대；대비H9아형적N2아형AIV진행확증，칙부출현1개특이성확증조대，즉341 bp N2기인조대；대비H9혹N2아형AIV화기타금호흡도병원체진행PCR확증，결과균위음성；결과표명해쌍중RT‐PCR최저능검출100 fg H9N2아형AIV적cDNA모판。
A duplex reverse transcription‐polymerase chain reaction(duplex RT‐PCR)was developed to de‐tect H9 and N2 subtype avian influenza viruses(AIVs)simultaneously .Two pairs of specific primers were designed according to the conserved regions on the sequences of H9 AIV‐HA gene and N2 AIV‐NA gene in GenBank .It was shown that all samples containing H9N2 subtype AIV could be amplified into two specific bands ,545 bp for H9 subtype AIV and 341 bp for N2 subtype AIV by this duplex RT‐PCR .All samples containing N2 subtype AIV with different HA genes (not H9) could be amplified into one specific band , 341 bp for N2 subtype AIV .No specific bands of the same sizes were amplified from other subtypes of AIVs and other avian pathogenic virus .As little as 100 fg of H9N2 subtype AIV could be detected by this duplex RT‐PCR .
