摘要
根据已发表的丝状霉形体簇各成员核苷酸序列,设计合成了2对引物McF、McR和MmcF、MmcR,建立了可以鉴别丝状霉形体山羊亚种(Mycoplasma mycoidessubsp.capri,Mmc)的巢式PCR方法。特异性和敏感性试验结果显示,该方法只能对Mmc扩增出195 bp的片段,而对其他病原菌不能扩增出任何条带,它最低能够检测出10 pg的Mmc DNA,说明该方法具有良好的特异性和很高的敏感性。田间试验结果显示,对4株霉形体分离株及其病料均可扩增出Mmc特异性片段,表明,该巢式PCR方法可用于Mmc快速鉴定和流行病学调查。
A nested polymerase chain reaction （PCR） assay for identifying M. rnycoides subsp, capri （Mmc） was developed. Two pairs of primers were designed and synthesized. The primers were designed according to nucleotide sequences of members of Mycoplasma mycoides cluster in the literature. Results showed that specificity and sensitivity of the assay may reach 10 pg of Mine DNA and a 195 bp-fragment from the strains PG3 and C87-1. No products from other pathogens were amplified. The 195 bp fragment was directly amplified from 4 Mycoplasma isolates, pathological lungs and pleural fluid by the nested PCR assay, respectively. The nested PCR assay could be used for rapid identification and epidemiological investigation of Mmc.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第11期940-943,共4页
Chinese Veterinary Science
基金
甘肃省农业生物技术专项(GNSW-2005-16)
关键词
丝状霉形体山羊亚种
巢式PCR
山羊传染性胸膜肺炎
Mycoplasma mycoides subsp, capri （Mmc）
nested PCR
contagious caprine pleuropneu-monia（CCPP）
