摘要：从河南某肉种鸡场疑似鸡滑液囊支原体感染的病鸡跗关节样品进行病原的分离与鉴定。分离菌经瑞氏染色,在油镜下呈球形或椭圆形。在固体培养基上表现为细小、光滑、致密的小菌落,菌落形态为"煎蛋状"。L型细菌鉴定发现菌体仍保持原有形态。菌体能够吸附红细胞,分解葡萄糖,但不能分解精氨酸和尿素,还可致SPF鸡胚死亡,说明该分离菌为支原体。进一步的血清学试验结果表明,该分离菌为鸡滑液囊支原体,而不是鸡毒支原体。根据已发表的鸡滑液囊支原体血凝素基因序列vlhA设计合成一对引物,经PCR扩增,该菌体能够扩增出鸡滑液囊支原体(773bp)的特异性片段。人工感染试验结果显示,SPF鸡能够复制出自然病例。

摘要：以HPS的16SrRNA中的基因序列设计合成引物,进行PCR扩增,标准菌株NR4、SW124、SW114、SW140和分离株XX、FS在预计的821bp位置上扩增出特异性条带,NJING株、胸膜肺炎(APP)等没有特异性条带产生。这证明PCR检测副猪嗜血杆菌具有较高的特异性和敏感性。

摘要：肺炎支原体的培养和保存条件非常苛刻,是疫苗规模化生产的工艺难题。本文针对猪支原体肺炎活疫苗(168株)生产工艺关键技术进行研究。通过培养试验筛选四种培养基配方表明该疫苗株在低血清改良培养基中生长良好;优化发酵培养工艺,使其在发酵罐培养60～70 h的峰值可达到1010CCU/mL;设计筛选该疫苗耐热保护剂和冻干工艺,37℃下保存10 d的耐老化试验结果显示,下降滴度小于100.5CCU/mL。本研究为提供高效、安全和稳定的猪肺炎支原体疫苗产品奠定基础。

摘要：[Objective] This study aimed to develop a quantitative competitive assay for detecting Mycoplasma hyopneumoniae in culture. [Method] One pair of Mhp-specific primers was designed for detecting Mhp in culture. Another pair of primers was designed based on the conserved gene sequences of Mycoplasma. The competitive template, which carried the same primer binding site with the target fragment, was constructed using enzyme digestion method. [Result] The logarithm of concentration of competitive template was treated as the abscissa（X-axis）, and the logarithm of corrected optical density ratio between amplification products by competitive template and target template was treated as the ordinate（Y-axis）. Thus the standard curve was drawn, and the regression equation was also obtained. When Y was assigned as 0, the concentration of the competitive plate was calculated, and then the concentration of Mhp was deduced. The logarithms of Color change unit（CCU） were treated as the abscissa（X-axis）, and the copy numbers of Mhp were treated as the ordinate（Y-axis）, so the standard curve was generated. It was found that the copy number of Mhp was highly correlated to CCU. [Conclusion] A quantitative competitive PCR assay was successfully established for the rapid detection of Mhp in culture.

摘要：根据猪肺炎支原体(Mhp)和猪鼻支原体(Mhr)的16S rRNA基因设计3条引物,建立Mhp和Mhr的双重PCR检测方法,并对该方法进行了特异性和敏感性试验。应用建立的方法检测了临床样品和疫苗样品。结果显示,该方法具有良好的特异性,最低可检测到0.66 ng的Mhp基因组DNA和0.58 ng Mhr基因组DNA。临床样品和疫苗样品检测结果与普通PCR检测结果一致。该双重PCR方法可用于Mhp与Mhr的鉴别、诊断以及疫苗纯粹性检查,快速而准确。