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鸡毒支原体PCR检测试剂盒的研制与应用
引用本文:谢芝勋,邓显文,唐小飞,谢志勤,庞耀珊,廖敏,刘加波,何竞铭.鸡毒支原体PCR检测试剂盒的研制与应用[J].畜牧与兽医,2004,36(1):3-5.
作者姓名:谢芝勋  邓显文  唐小飞  谢志勤  庞耀珊  廖敏  刘加波  何竞铭
作者单位:广西兽医研究所,广西,南宁,530001
基金项目:广西科技厅攻关项目 (991 1 0 4 3)
摘    要:根据基因库中鸡毒支原体 1 6SrRNA的序列研制PCR检测试剂盒 ,用于检测鸡毒支原体 (MG)。结果表明该MG PCR检测试剂盒对不同MG参考菌株和地方分离株均能特异性地扩增出 732bp条带 ,而对其他禽支原体和禽病病原体的扩增结果为阴性。该MG PCR试剂盒最低能检测出 1 0 0fg的MGDNA模板。保存期测定结果表明 ,该MG PCR试剂盒在 - 2 0℃条件下保存至 1 ,3 ,6和 9个月时 ,其敏感性无明显变化 ,仍能检测到 1 0 0fg至 1pg的MGDNA模板。保存至 1 2个月时其敏感度虽降低了 1个滴度 ,但仍能 1 0 0 %检出人工感染鸡的临床样品

关 键 词:鸡毒支原体  聚合酶链反应  试剂盒
文章编号:0529-5130(2004)01-0003-03
修稿时间:2003年6月27日

The studies and application of PCR kit for Mycoplasma gallisepitcum detection
XIE Zhi-xun,DENG Xian-wen,TANG Xiao-fei,XIE Zhi-qin,PANG Yao-shang,LIAO Min,LIU Jia-bo,HE Jing-ming.The studies and application of PCR kit for Mycoplasma gallisepitcum detection[J].Animal Husbandry & Veterinary Medicine,2004,36(1):3-5.
Authors:XIE Zhi-xun  DENG Xian-wen  TANG Xiao-fei  XIE Zhi-qin  PANG Yao-shang  LIAO Min  LIU Jia-bo  HE Jing-ming
Abstract:A PCR kit was developed for Mycoplasma gallisepitcum (MG) detection. The PCR primers based on the 16S rRNA sequence of MG could amplified a specific 732 bp DNA product by the PCR kit from MG reference strains and isolates strains,but no specific band was amplified from other avian mycoplasma(including MS,MI and MM) and other avian pathogenic viruses and bacteria.As little as 100fg MG DNA was detected using gel electrophonesis in this PCR kit. After stored at -20℃ for 1,3,6 and 9 months respectively, the kits still could be ued to detect from 100 fg to 1 pg MG DNA.Although the sensitivity of the kit is 10 fold lower after stored for 12 months,100% of the clinical samples of infected chickens could be detected positively with the MG-PCR kit.
Keywords:Mycoplasma gallisepitcum  polymerase chain reaction  kit
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