| 鸡传染性贫血病毒套式PCR检测方法的建立 |
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| 引用本文： | 王景儒,杨晓静.鸡传染性贫血病毒套式PCR检测方法的建立[J].中国畜牧兽医,2013,40(4):55-58. |
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| 作者姓名： | 王景儒 杨晓静 |
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| 作者单位： | 辽宁朝阳工程技术学校,辽宁朝阳 122000 |
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| 摘 要： | 本试验根据GenBank中登录的禽传染性贫血病毒(CAV)基因序列,设计合成2对引物,外引物的扩增片段大小为485 bp,内引物的扩增片段大小为297 bp,建立了适合CAV快速检测的套式PCR方法(nested PCR),并采用该方法对CAV阳性毒株及临床病料进行了检测。结果显示,该方法能扩增到297 bp的条带,禽流感病毒(H9亚型)、新城疫病毒、传染性法氏囊病病毒、禽网状内皮增生病病毒、减蛋综合征病毒、禽呼肠孤病毒、马立克氏病病毒的扩增结果均为阴性。该方法第1步扩增的敏感性是100 pg,第2步PCR扩增的敏感性是1 fg,敏感性提高了105倍。本研究建立的CAV套式PCR方法具有敏感性高、重复性好、特异性强等优点,可用于CAV的临床诊断和分子流行病学调查等。
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| 关 键 词： | 禽传染性贫血病病毒 套式PCR 检测 |
| 收稿时间： | 2012-09-21 |
Establishment of a Nested PCR Assay for Detection of Chicken Infectious Anemia Virus |
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| WANG Jing-ru
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YANG Xiao-jing.Establishment of a Nested PCR Assay for Detection of Chicken Infectious Anemia Virus[J].China Animal Husbandry & Veterinary Medicine,2013,40(4):55-58. |
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| Authors: | WANG Jing-ru YANG Xiao-jing |
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| Institution: | Liaoning Chaoyang Engineering Technical School,Chaoyang 122000,China |
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| Abstract: | According to the sequence of chicken infectious anemia virus (CIAV) strain published in GenBank,two pairs of primers were designed and synthesized.The outer primers amplified a fragment of 485 bp in length, and the inner primers amplification fragment size was 297 bp in length.A nested PCR assay for rapid detection of ALV was established.A specific 297 bp fragment was amplified from DNA templates of CAV strain,but no bands were amplified with templates extracted respectively from avian influenza virus (AIV) subtype H9,Newcastle disease virus (NDV), infectious bursal disease virus(IBDV),egg drop syndrome virus (EDSV), reticuloendotheliosis (REV), avian reovirus (ARV) and Marek’s disease virus(MDV). Sensitivity of the 1st and 2nd amplifications by the nested PCR assay were 100 pg and 1 fg,respectively.The sensitivite of the 2nd amplifications increased by 105 times.The results showed that the nested PCR was specific,sensitive,rapid,and accurate,and could be used as a routine assay for the detection of CAV.This method had good reproducibility, specificity and sensitivity, and might detect low content CAV accurately and rapidly.This method could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of CAV. |
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| Keywords: | chicken infectious anemia virus nested PCR detection |
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