PCR检测鸡毒霉形体和鸡滑液囊霉形体的研究

根据禽霉形体 16SrRNA的基因序列设计、合成了 1对引物 ,用这对引物对鸡毒霉形体 (Mycoplasmagallisep ticum ,MG)和鸡滑液囊霉形体 (Mycoplasmasynoviae ,MS)菌株DNA进行PCR扩增 ,得到了与预期大小相一致的约 5 80bp的PCR产物 ,而这对引物对其他禽病病原DNA或RNA模板的扩增结果为阴性。PCR方法对MG和MS的最小检出量分别为 2 pg和 3pg。应用已建立的PCR方法检测MG人工感染样品和临床样品 ,从人工感染样品中均检测到MG ,临床样品的MG阳性检出率为 10 .2 5 % ,高于常规分离培养的阳性检出率

Detection of Mycoplasma gallisepticum and M. synoviae by polymerase chain reaction

A single set of oligonucleotide primers was designed based on 16Sribosomal RNA (rRNA) sequences of avian Mycoplasma. This set of primers selectively amplifies a 580bp product from Mycoplasma gallisepticum (MG) and M. synoviae (MS), but does not amplify the DNA or RNA of other bacteria or viruses. The detection threshold of polymerase chain reaction (PCR) of MGand MSwas 2pg or 3pg. The method had been successfully used to detect MG or MS in different samples. The preliminary results showed that this technique can detect MG fromallof the experimentally infected samples. Among them, the positive detection rate in throat swabs and trachea samples was the highest. Using PCR, 10.25% of field samples was positively detected, it was higher than that of the routine culture. It indicated that this technique is a useful screening assay for the detection of M.gallisepticum and M.synoviae.