| 使用免疫BMPs纯化细菌PCR反应模板的比较 |
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| 引用本文： | 田菊霞,解宇,杨艳宏.使用免疫BMPs纯化细菌PCR反应模板的比较[J].科技通报,2012,28(5):80-83. |
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| 作者姓名： | 田菊霞 解宇 杨艳宏 |
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| 作者单位： | 杭州师范大学,杭州,310036 |
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| 基金项目： | 杭师大重大重点预研项目，杭州市科技发展项目，浙江省科技计划项目 |
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| 摘 要： | 目的:使用免疫BMPs,对含有大肠杆菌O157的粪便标本,进行目标病原菌的捕获和洗净,建立一种快速纯化细菌PCR反应模板的方法。方法:将大肠杆菌O157抗体接种于BMPs表面,制成免疫BMPs。使用免疫BMPs,对含有不同浓度大肠杆菌O157的粪便标本,进行目标病原菌的捕获、分离和洗净,再使用洗净的病原菌制备PCR反应模板。作为对照,对含有不同浓度大肠杆菌O157的粪便标本,常规进行增菌培养及鉴别培养后,制备细菌的PCR反应模板,或粪便标本直接制备PCR反应模板。结果:使用免疫BMPs处理后制备PCR反应模板,与通过常规增菌培养和鉴别培养后制备PCR反应模板,PCR检测结果一致。而粪便标本直接取样制备PCR反应模板,大肠杆菌O157低浓度时,PCR检测的阳性率较低。结论:利用免疫BMPs捕获自然标本(粪便)中的目标病原菌,通过分离和洗净,去除标本中存在的PCR反应抑制物等影响因素,可省略增菌培养及鉴别培养,快速纯化目标病原菌的PCR反应模板。
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| 关 键 词： | 免疫BMPs PCR反应模板 纯化 |
Comparison of PCR Reaction Templates for Purifying Bacteria with Immune BMPs |
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| TIAN Juxia
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XIE Yu
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YANG Yanhong.Comparison of PCR Reaction Templates for Purifying Bacteria with Immune BMPs[J].Bulletin of Science and Technology,2012,28(5):80-83. |
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| Authors: | TIAN Juxia XIE Yu YANG Yanhong |
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| Institution: | (Medical College of Hangzhou Normal University,Hangzhou 310036,China) |
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| Abstract: | Objective To Use the immune BMPs for capturing and purifying target pathogen in stool specimens which contain E.coli O157 and establish a rapid PCR reaction to purify the bacteria.Methods Inoculating E.coli O157 antibodies to the surface of BMPs for making immune BMPs,and using the immune BMPs to capture,separate and purify target pathogen in stool specimens which contain different concentrations of E.coli O157,and then using the purified pathogens as PCR reaction template.In comparison,stool specimens containing different concentrations of E.coli O157 were enrichedly cultured or differentially cultured and then were made for the PCR reaction template,or the fecal samples were directly prepared for the PCR reaction template.Results The PCR reaction template from the immune BMPs treatment and the comparison templates through the culture treatments have the identical PCR test results.While the template prepared directly from fecal specimens is low in concentration of E.coli O157 and low in PCR positive rate of detection.Conclusion Using immune BMPs for capturing the target pathogens in the natural samples(faeces),as well as separating and purifying the inhibiting factors to PCR reaction existing in the specimens,can omit the enriched culture and differential culture process and rapidly purify the PCR reaction template of target pathogens. |
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| Keywords: | Immune Bacterial Magnetic Particles(Immune BMPs) PCR reaction template Purification |
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