摘要
根据Ⅰ群禽腺病毒(fowl adenovirusⅠ,FAVⅠ)六邻体(hexon)基因序列,设计合成2对引物,建立了适合FAVⅠ快速检测的套式PCR方法(nested PCR)。应用该方法对FAVⅠ阳性毒株及临床病料进行检测,2步PCR扩增片段大小分别为475和237bp,而其他7种禽源病毒的扩增结果均为阴性;该套式PCR方法扩增的敏感性为100pg和1fg,敏感性提高了105倍;所建立的FAVⅠ套式PCR方法大大提高了常规PCR的特异性和敏感性,重复性好,可以检测出极低含量的FAVⅠ,可用于FAVⅠ的临床诊断和分子流行病学调查等。
According to the sequence of hexon gene of fowl adenovirus group I （FAV T ） strain published in Gen]3ank, two pairs of primers were designed and synthesized. The outer primers amplified a fragment of 475 bp in length, and the inner primers amplification fragment was 237 bp in length. A nested PCR assay for rapid detection of FAV I was established. A spe- cific 237 bp fragment was amplified from DNA templates of FAV I strain,but no bands were amplified with templates extrac- ted respectively from avian influenza virus （AIV） subtype H9,Newcastle disease virus （NDV）, infectious bursal disease virus （IBDV） ,duck plague virus （DPV）, reticuloendotheliosis （REV）, avian reovirus （ARV）, Marek^s disease virus （MDV）. Sen- sitivity 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,accurate,and could be used as a routine assay for the detection of FAV I This method had good reproducibility, specificity and sensitivity, and might detect low content FAV I accurately and rapidly. This method could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of FAV ~[.
出处
《中国畜牧兽医》
CAS
北大核心
2013年第8期30-33,共4页
China Animal Husbandry & Veterinary Medicine
