应用多重聚合酶链反应检测鸡毒霉形体和滑液霉形体的研究

根据鸡毒霉形体（ ＭＧ） 、滑液霉形体（ ＭＳ） 的基因文库，设计并合成了分别与ＭＧ、ＭＳ 某段基因序列互补的两对引物，用这两对引物进行多重聚合酶链反应（ ＭｕｌｔｉＰＣＲ） 同时检测ＭＧ 与ＭＳ。当样品中同时含有ＭＧ 和 ＭＳ 的目的模板ＤＮＡ 时，均同时得到２ 条大小与试验设计相符的７３２ ｂｐ（ ＭＧ） 和２０７ ｂｐ （ ＭＳ） 的ＰＣＲ 扩增带；而对其它８ 种禽病病原的扩增均为阴性。敏感性测定结果表明，该多重ＰＣＲ 技术最低能同时检出１００ ｆｇ 的ＭＧ、ＭＳ ＤＮＡ 模板量

Establishment of a Multi polymerase Chain Reaction (multi PCR) for Detecting Mycoplasma gallisepticum and Mycoplasma synoviae

On the basis of the gene library bank of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) the two pairs of primers that are complementary to a given gene sequence of MG and MS were designed and synthesized, with which a multi PCR was performed for detecting MG and MS, when the objective template DNAs of MG and MS existed in samples the PCR amplified two bands the sizes of which are as same as bands experimentally designed as one 732 bp (MG) and as another one 207 bp (MS) would be obtained. However, amplification of other different eight pathogens of avian diseases all were negative. The sensitivity test revealed that 100 fg of DNA of MG and MS can be detected with the multi PCR.