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目的:建立1种快速检测鸡肉中布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的多重PCR方法。方法:根据3种弓形菌的rpoB基因设计了4条引物,进行多重PCR扩增,测试PCR体系的特异性和灵敏度,并应用于鸡肉样品的检测。结果:对布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的基因组DNA能够特异性扩增出373,149和236 bp大小的目的条带,检测灵敏度分别为1,10,10 pg。16份鸡肉样品中,7份样品被检出布氏弓形菌,3份样品被检出嗜低温弓形菌,1份样品被检出斯氏弓形菌,与常规分离培养法检测的符合率分别为100%,87.5%和100%。结论:建立的多重PCR法具有操作简便,检测周期短,特异性强和灵敏度高的特点,能够实现对鸡肉中布氏弓形菌、嗜低温弓形菌和斯氏弓形菌的同时快速检测和监控。
OBJECTIVE: To establish a multiplex PCR method for rapid detection of toxoplasma & lt; RTI ID = 0.0 & gt; Broth, & lt; / RTI & gt; Methods: Four primers were designed according to rpoB gene of three kinds of Toxoplasma gondii, amplified by multiplex PCR, tested the specificity and sensitivity of PCR system and applied to the detection of chicken samples. Results: The target bands of 373, 149 and 236 bp were amplified specifically from T. gibberei, T. hyodysenteriae and T. histotans. The detection sensitivity was 1, 10 and 10 pg, respectively. Among 16 chicken samples, 7 samples were identified as T. paratyphi, 3 samples were tested for T. hyphae and 1 sample was tested for T. tomentosa. The coincidence rates with routine isolation and culture methods were 100%, 87.5% and 100%. Conclusion: The established multiplex PCR method has the advantages of simple operation, short detection cycle, strong specificity and high sensitivity, and can simultaneously detect and monitor the toxoplasma & amp;