国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2009年
3期
144-146
,共3页
袁忠英%沈玉娟%曹建平%周何军%卢潍媛%陈勤%倪奕昌%汤林华
袁忠英%瀋玉娟%曹建平%週何軍%盧濰媛%陳勤%倪奕昌%湯林華
원충영%침옥연%조건평%주하군%로유원%진근%예혁창%탕림화
环介导等温扩增法%牛隐孢子虫%18SrRNA
環介導等溫擴增法%牛隱孢子蟲%18SrRNA
배개도등온확증법%우은포자충%18SrRNA
Loop-mediated isothermal amplification%Cryptosporidium bovis%18S rRNA
目的 用环介导等温扩增技术检测牛源隐孢子虫.方法 提取牛粪中隐孢子虫卵囊DNA.根据隐孢子虫18S rRNA序列及环介导等温扩增(100p-mediated isothermal amplification,LAMP)技术的原理,设计4条隐孢子虫特异引物,利用LAMP检测牛粪中隐孢子虫卵囊DNA和蓝氏贾第鞭毛虫DNA,以正常牛粪DNA为阴性对照.LAMP产物经SYBR Green I显色及电泳后观察结果,绿色判为阳性,棕色判为阴性,对LAMP产物进行电泳分析,观察其特征条带的情况.结果隐孢子虫卵囊DNA检测管经显色后时不再呈绿色,阴性对照及蓝氏贾第鞭毛虫DNA呈棕色.隐孢子虫LAMP产物经电泳后呈LAMP特征性梯状条带,阴性对照及蓝氏贾第鞭毛虫DNA无扩增产物.结论 LAMP方法敏感、特异、简便,可用于检测牛粪中的隐孢子虫.
目的 用環介導等溫擴增技術檢測牛源隱孢子蟲.方法 提取牛糞中隱孢子蟲卵囊DNA.根據隱孢子蟲18S rRNA序列及環介導等溫擴增(100p-mediated isothermal amplification,LAMP)技術的原理,設計4條隱孢子蟲特異引物,利用LAMP檢測牛糞中隱孢子蟲卵囊DNA和藍氏賈第鞭毛蟲DNA,以正常牛糞DNA為陰性對照.LAMP產物經SYBR Green I顯色及電泳後觀察結果,綠色判為暘性,棕色判為陰性,對LAMP產物進行電泳分析,觀察其特徵條帶的情況.結果隱孢子蟲卵囊DNA檢測管經顯色後時不再呈綠色,陰性對照及藍氏賈第鞭毛蟲DNA呈棕色.隱孢子蟲LAMP產物經電泳後呈LAMP特徵性梯狀條帶,陰性對照及藍氏賈第鞭毛蟲DNA無擴增產物.結論 LAMP方法敏感、特異、簡便,可用于檢測牛糞中的隱孢子蟲.
목적 용배개도등온확증기술검측우원은포자충.방법 제취우분중은포자충란낭DNA.근거은포자충18S rRNA서렬급배개도등온확증(100p-mediated isothermal amplification,LAMP)기술적원리,설계4조은포자충특이인물,이용LAMP검측우분중은포자충란낭DNA화람씨가제편모충DNA,이정상우분DNA위음성대조.LAMP산물경SYBR Green I현색급전영후관찰결과,록색판위양성,종색판위음성,대LAMP산물진행전영분석,관찰기특정조대적정황.결과은포자충란낭DNA검측관경현색후시불재정록색,음성대조급람씨가제편모충DNA정종색.은포자충LAMP산물경전영후정LAMP특정성제상조대,음성대조급람씨가제편모충DNA무확증산물.결론 LAMP방법민감、특이、간편,가용우검측우분중적은포자충.
Objective To detect calf-origin Cryptosporidium bovis by loop-mediated isothermal amplifi-cation(LAMP). Methods The DNA was extracted from calf faeces infected with Cryptosporidium.Accord-ing to Cryptosporidium 18S rRNA sequence and the mechanism of LAMP,4 Cryptosporidium specific primers were designed for LAMP detection of Cryptosporidium and Giardia lamblia.the DNA from normal calf faeces and Giardia lamblia were used for the negative contr01.The LAMP products were stained by SYBR Gteen I and analyzed by eleetrophoresis. Results After staining.the LAMP products from Cryptosporidium oocysts DNA turned green while the negative control and Giardia lamblia DNA kept brown.Electrophoresis analysis showed the LAMP product of Cr379tosporidium Was with characteristic ladders.but products of the negative control and Giardia lamblia Was not. Conclusion LAMP assay iS a simple,sensitive and specific method for detection Cryptosporidium in cattle faeces.
