| 用PCR和探针杂交法检测鸡败血霉形体 |
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| 引用本文： | 任家琰,霍乃蕊,郭建华.用PCR和探针杂交法检测鸡败血霉形体[J].中国兽医学报,2000,20(2):156-159. |
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| 作者姓名： | 任家琰 霍乃蕊 郭建华 |
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| 作者单位： | 山西农业大学动物医学系!山西太谷030801 |
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| 基金项目： | 山西省自然科学基金资助项目! ( 9710 68) |
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| 摘 要： | 根据鸡败血霉形体ｆＭＧ－２核酸片断序列，设计合成了１对２５ｂｐ寡核苷酸引物，对鸡败血霉形体基因组ＤＮＡ进行扩增，均获得预期的７３２ｂｐ扩增产物，检测灵敏度为１ｂｐ；参考菌株ＤＮＡ无扩增。回收纯化琼脂糖电泳凝胶中的扩增产物，ＤＩＧ随机引物法合成核酸探讨，Ｄｏｔ－ｂｌｏｔ杂交试验，鸡败血霉形体呈阳性，检测灵敏度为１００ｐｇ；其他为阴性。对自然发病鸡群检测进一步表明，建立的ＰＣＲ和探针杂交法具有高度的灵
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| 关 键 词： | PCR 探针杂交 鸡败血霉形体 检测 |
Detection of Mycoplasma gallisepticum by PCR and DNA probe hybridzation |
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| REN Jia-yan,HOU Nai-rui,GUO Jian-hua.Detection of Mycoplasma gallisepticum by PCR and DNA probe hybridzation[J].Chinese Journal of Veterinary Science,2000,20(2):156-159. |
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| Authors: | REN Jia-yan HOU Nai-rui GUO Jian-hua |
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| Abstract: | A pair of 25 base primers were designed and synthesized according to the nucleotide fragment(fMG 2) sequence of Mycoplasma gallisepticum. The primers were applied to amplify Mycoplasma gallisepticum DNA by PCR and expected amplification of a 732 bp product was demonstrated by ethidium bromide staining of 1.5% agarose gel electrophoresis,its sensitivity to be 1 pg without the control strain DNA to be amplified.Thereafter the PCR product was used as a probe labelled with DIG 11 dUTP via random primer labelling to hybridize with the DNA above. The Dot blot assay suggested that the probe hybridized with DNA of Mycoplasma gallisepticum but not with that of negative control strains,its sensitivity to be 100 pg.The further detection to naturally infected chickens showed that the two methods developed in this study had a high sensitivity and specificity,which could be applied to detection of early and latent infection of Mycoplasma gallisepticum. |
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| Keywords: | PCR probe hybridization Mycoplasma gallisepticum |
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