|
|
|
丝状霉形体山羊亚种巢式PCR检测方法的建立
|
|
|
|
储岳峰， 逯忠新， 赵萍， 高鹏程， 贺英
|
|
|
|
（中国农业科学院 兰州兽医研究所 家畜疫病病原生物学国家重点实验室 农业部畜禽病毒学
重点开放实验室，甘肃 兰州730046）
|
|
|
|
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 mycoides subsp. 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195bp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10pg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。 |
|
关键词：
|
|
丝状霉形体山羊亚种；巢式PCR；山羊传染性胸膜肺炎 |
|
中图分类号：S 852.62：Q 503 文献标识码：
A 文章编号：1673-4696(2007)11-0940-04 |
|
|
Development of nested PCR assay for detecting Mycoplasma
mycoides subsp. capri |
|
|
|
CHU Yue-feng,LU Zhong-xin,ZHAO Ping,GAO Peng-cheng,HE Ying
|
|
|
|
(Key Laboratory of Animal Virology of the Ministry of Agriculture/State Key Laboratory of Veterinary Etiological
Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
|
|
Abstract: A nested polymerase chain reaction (PCR) assay for identifying M. mycoides 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 10pg of Mmc DNA and a 195bp-fragment from the strains PG3 and C87-1. No products from other pathogens were amplified. The 195bp 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. |
|
|
|
Key words: Mycoplasma mycoides subsp. capri (Mmc);nested PCR;contagious caprine pleuropneumonia(CCPP) |
|
|
|
|
|
《中国兽医科学》 ©
版权所有
Chinese Veterinary Science
zgsykx@zgsykx.com |
