CAJ | 학술논문

马立克氏病（MD）是养禽业最重要的疫病之一，一直缺少有效的早期诊断方法。根据血清I型马立克氏病毒（MDV1）meq基因的核酸序列设计了一对寡苷酸核引物，分别对MDV1致瘤株（京－1株）、非致瘤株（MD11／75C株、CV1988株）、MDV2（SB-1株）、HVT（Fc-126株)的核酸进行扩增。结果表明：京－1株扩增到约1.15kb核酸片段，MD11／75C株和CVI988株扩增到约1.0kb核酸片段，而SB-1株、Fc-126株没得到任何扩增产物。PCR产物Dotblot结果显示，京－1株、MD11／75C株和CVI988株的扩增产物都与Digoxigenin标记的meq基因探针杂交，说明都是特异性的扩增产物。对MSB1细胞DNA及MDV感染鸡的血液及肝、肾肿瘤等DNA扩增都得到1.15kb条带。将京－1株和CVI988株感染的细胞DNA混合再扩增，同时得到1.15kb和1.0kb的核酸条带，所以根据扩增产物大小可以区别致瘤株京－1株及非致瘤株CV1988株，这表示可从CVI988株病毒免疫鸡体内检测到MDV强毒，适于早期确诊强毒感染。
마립극씨병（MD）시양금업최중요적역병지일，일직결소유효적조기진단방법。근거혈청I형마립극씨병독（MDV1）meq기인적핵산서렬설계료일대과감산핵인물，분별대MDV1치류주（경－1주）、비치류주（MD11／75C주、CV1988주）、MDV2（SB-1주）、HVT（Fc-126주)적핵산진행확증。결과표명：경－1주확증도약1.15kb핵산편단，MD11／75C주화CVI988주확증도약1.0kb핵산편단，이SB-1주、Fc-126주몰득도임하확증산물。PCR산물Dotblot결과현시，경－1주、MD11／75C주화CVI988주적확증산물도여Digoxigenin표기적meq기인탐침잡교，설명도시특이성적확증산물。대MSB1세포DNA급MDV감염계적혈액급간、신종류등DNA확증도득도1.15kb조대。장경－1주화CVI988주감염적세포DNA혼합재확증，동시득도1.15kb화1.0kb적핵산조대，소이근거확증산물대소가이구별치류주경－1주급비치류주CV1988주，저표시가종CVI988주병독면역계체내검측도MDV강독，괄우조기학진강독감염。
Marek's disease is one of the most important infectious diseasesin poultry farm, but still there are lack of effective methods for diagnosing it. A polymerase chain reaction(PCR) assay based on primers flanking meq gene of oncogenic MDV1 (J-l strain) was developed. These primers amplified a 1.15kb nucleotide band from oncogenic strain J-1 and a 1.0kb band from non-oncogenic strains(MD11/75c, CVI988). The MDV1 specific primers didn't amplify MDV2(SB-1) and HVT(Fc-126) DNA. Dot blot of the PCR products showed that the products of strains J-1,MD11/75c and CVI988 could hybridize with meq gene probe (labeled by digoxigenin). Mixing the J-1 DNA with CVI988 DNA as template, the results indicated the primers could amplify the two bands(1.15kb and 1.0kb) from the mixture, these primers can also amplify meq gene from MSB1 cell line and also from blood and tumor samples of J-l infected chickens. Therefore, using this primer, we can distinguish between oncogenic and non-oncogenic strain of MDV1.