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聚合酶链反应检测绵羊肺炎霉形体的研究

张莉1， 杨壮省2， 王英珍1， 鄢明华1， 孙继国3

（1. 天津市畜牧兽医研究所， 天津031120； 2. 邢台市畜牧局， 河北 邢台054001；
3. 河北农业大学， 河北 保定071001）

摘要： 参照基因库中已发表的绵羊肺炎霉形体Y98株的16S rDNA序列，设计合成了1对引物，建立了聚合酶链反应(PCR)检测绵羊肺炎霉形体的方法。结果显示，所建立的PCR能特异扩增绵羊肺炎霉形体的DNA，而对照的菌株均为阴性；其敏感性可达1pg。经对绵羊肺炎霉形体分离株HD1的扩增产物进行测序，并与绵羊肺炎霉形体标准株Y98的16S rDNA序列进行比较，发现有6个碱基的差异。

关键词： 绵羊肺炎霉形体； 聚合酶链反应； 16S rDNA

中图分类号：S 852.62  文献标识码： A  文章编号：1000-6419(2004)11-0007-04

Development of polymerase chain reaction for detection 
of Mycoplasma ovipneumoniae in sheep

ZHANG Li1, YANG Zhuangsheng2, WANG Yingzhen1, 
YAN Minghua1, SUN Jiguo3

(1. Tianjin Institute of Animal Science and Veterinary Medicine, Tianjin 031120,China;
2. Hebei Bureau of Animal Husbandry, Xingtai 054001,China;
3. Hebei Agricultural University, Baoding 071001,China)

Abstract: A PCR method was developed with two oligonucleotide primers(ZW5 and ZW3 ), which were designed on the base of 16S rDNA gene of M.ovipneumoniae standard strain Y98. Only a target DNA fragment was amplified from M.ovipneumoniae〖KG*2〗DNA by the PCR. The sensitivity〖KG*2〗of the PCR technique was 1pg.The amplified HD1 strain DNA fragment was compared with 16S rDNA part sequence of Y98 strain postsequencing. The results showed that these were six base’ differences between them.

Key words: Mycoplasma ovipneumoniae; microbiology; polymerase chain reaction (PCR)

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