| 地高辛素、生物素标记产毒性大肠杆菌 Ⅰ型不耐热肠毒素ADN探针的制备及应用 |
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| 引用本文： | 曹纬,彭文伟,谢彦博,余守义,黄君健,陈忠义. 地高辛素、生物素标记产毒性大肠杆菌 Ⅰ型不耐热肠毒素ADN探针的制备及应用[J]. 中山大学学报(医学科学版), 1990, 0(1) |
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| 作者姓名： | 曹纬 彭文伟 谢彦博 余守义 黄君健 陈忠义 |
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| 作者单位： | 中山医科大学传染病学教研室，中山医科大学传染病学教研室，中山医科大学传染病学教研室，第一军医大学，第一军医大学，第一军医大学 研究生，江苏省南通医学传染病学教研室 |
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| 摘 要： | 采用随机引物法制备地高辛素、生物素和~(32)P标记产毒性大肠杆菌Ⅰ型不耐热肠毒素(LT_1)DNA探针。地高辛素、生物素和~(32)P LT_1探针分别在DNA斑点杂交试验中检测到0.1pg、1pg和0.1pg的质粒DNA,在菌落原位杂交试验中检测到12、120和12个细菌/ml,在粪便斑点杂交试验中检测到24、240和24个细菌/ml。采用RNA酶A和蛋白酶K消化处理滤膜,消除了假阳性,使得地高辛素、生物素LT_1探针的特异性与~(32)PLT_1探针完全一致。检测143株自腹泻病人分离的大肠杆菌,14株(9.79％)阳性,3种探针结果相同。
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| 关 键 词： | 地高辛素 生物素 探针 肠毒素 |
PREPARATION AND APPLICATION OF DIGOXIGENIN-LABELED AND BIOTINYLATED TYPE I HEAT-LABILE TOXIN |
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| Cao Wei Peng Wenwei Xie Yanbo. PREPARATION AND APPLICATION OF DIGOXIGENIN-LABELED AND BIOTINYLATED TYPE I HEAT-LABILE TOXIN[J]. Journal of Sun Yatsen University(Medical Sciences), 1990, 0(1) |
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| Authors: | Cao Wei Peng Wenwei Xie Yanbo |
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| Affiliation: | Cao Wei Peng Wenwei Xie Yanbo(Sun Yat-Sen University of Medical Sciences)Yu Shouyi Huang Junjlan Cheng Yizhong(The First Army Univesity of Medical Sciences) |
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| Abstract: | Type I heat-labile toxin(LT1) DNA probes were labeled with digoxigenin, biotin, and 32P by random priming. Digoxigenin labeled, biotinylated, and 32P LTj probe detected 0.1 Pg, 1 Pg. and 0.1 Pg of EWD299 DNA in DNA blot hybridization; 12, 120 and 12 bacteria per millilitre of normal saline in bacteria in situ hybridization; 24, 240, and 24 bacteria per millilitre of stool suspension in stool blot hybridization after exposure to indicator dyes for 4 hours or X-ray film for 72 hours. The treatment of nitrocellulose membrane filters with RNase A and proteinase K eliminated the possibility of false-positivity and made the specificity of digoxigenin labeled and biotinylated LT: probes as good as 32PLTI probe. 9.79% (14/143) Escherichia coli from diarrheal patients were detected by these three probes. |
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| Keywords: | Digoxigenin Biotin Probe Enterotoxin |
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