药物不良反应杂志
藥物不良反應雜誌
약물불량반응잡지
ADVERSE DRUG REACTIONS JOURNAL
2013年
1期
11-16
,共6页
龙鑫%孙承业%谢立璟%王英伟%刘冰
龍鑫%孫承業%謝立璟%王英偉%劉冰
룡흠%손승업%사립경%왕영위%류빙
有毒植物%随机扩增多态性DNA标记技术%简单重复序列间标记技术%DNA条形码技术
有毒植物%隨機擴增多態性DNA標記技術%簡單重複序列間標記技術%DNA條形碼技術
유독식물%수궤확증다태성DNA표기기술%간단중복서렬간표기기술%DNA조형마기술
Poisonous plants%Random amplified polymorphic DNA%Inter-simple sequences repeats%DNA barcoding
目的 评价3种不同的分子标记技术对植物属级的鉴定能力以选出最适合在中毒现场快速鉴定有毒植物的分子技术. 方法 选取毛茛科、大戟科共18种19份有毒植物叶片样品,用改良十六烷基三甲基溴化铵法提取基因组DNA,分别使用随机扩增多态性DNA标记(RAPD)、简单重复序列间标记(ISSR)和DNA条形码技术进行物种鉴定(属级),使用NYSTS、SPSS、PAUP、MEGA软件进行聚类分析,比较3种DNA分子标记技术的准确性、可靠性、时效性和可操作性. 结果 准确性:RAPD技术未能鉴定样品的属级,ISSR技术和DNA条形码技术的鉴定准确率分别为68％和100％.可靠性:RAPD技术与ISSR技术主观影响率分别为44％、26％,条带重复率分别为47％、45％；DNA条形码技术trnH-psbA片段扩增成功率与测序成功率均为100％.时效性:从总DNA提取开始至得到最终鉴定结果,RAPD、ISSR、DNA条形码技术耗时分别为8.5、9.0、42.2 h.可操作性:RAPD与ISSR技术的鉴定工作可在普通实验室完成,DNA条形码技术的鉴定工作则需要特殊的测序设备. 结论 初步认为DNA条形码技术是比较适用于突发有毒植物中毒事件中快速病因判定的分子鉴定技术.
目的 評價3種不同的分子標記技術對植物屬級的鑒定能力以選齣最適閤在中毒現場快速鑒定有毒植物的分子技術. 方法 選取毛茛科、大戟科共18種19份有毒植物葉片樣品,用改良十六烷基三甲基溴化銨法提取基因組DNA,分彆使用隨機擴增多態性DNA標記(RAPD)、簡單重複序列間標記(ISSR)和DNA條形碼技術進行物種鑒定(屬級),使用NYSTS、SPSS、PAUP、MEGA軟件進行聚類分析,比較3種DNA分子標記技術的準確性、可靠性、時效性和可操作性. 結果 準確性:RAPD技術未能鑒定樣品的屬級,ISSR技術和DNA條形碼技術的鑒定準確率分彆為68％和100％.可靠性:RAPD技術與ISSR技術主觀影響率分彆為44％、26％,條帶重複率分彆為47％、45％；DNA條形碼技術trnH-psbA片段擴增成功率與測序成功率均為100％.時效性:從總DNA提取開始至得到最終鑒定結果,RAPD、ISSR、DNA條形碼技術耗時分彆為8.5、9.0、42.2 h.可操作性:RAPD與ISSR技術的鑒定工作可在普通實驗室完成,DNA條形碼技術的鑒定工作則需要特殊的測序設備. 結論 初步認為DNA條形碼技術是比較適用于突髮有毒植物中毒事件中快速病因判定的分子鑒定技術.
목적 평개3충불동적분자표기기술대식물속급적감정능력이선출최괄합재중독현장쾌속감정유독식물적분자기술. 방법 선취모간과、대극과공18충19빈유독식물협편양품,용개량십륙완기삼갑기추화안법제취기인조DNA,분별사용수궤확증다태성DNA표기(RAPD)、간단중복서렬간표기(ISSR)화DNA조형마기술진행물충감정(속급),사용NYSTS、SPSS、PAUP、MEGA연건진행취류분석,비교3충DNA분자표기기술적준학성、가고성、시효성화가조작성. 결과 준학성:RAPD기술미능감정양품적속급,ISSR기술화DNA조형마기술적감정준학솔분별위68％화100％.가고성:RAPD기술여ISSR기술주관영향솔분별위44％、26％,조대중복솔분별위47％、45％；DNA조형마기술trnH-psbA편단확증성공솔여측서성공솔균위100％.시효성:종총DNA제취개시지득도최종감정결과,RAPD、ISSR、DNA조형마기술모시분별위8.5、9.0、42.2 h.가조작성:RAPD여ISSR기술적감정공작가재보통실험실완성,DNA조형마기술적감정공작칙수요특수적측서설비. 결론 초보인위DNA조형마기술시비교괄용우돌발유독식물중독사건중쾌속병인판정적분자감정기술.
Objective To evaluate the efficiency of the three molecular marker techniques for identification of plant genus in order to select the most suitable molecular technique to rapidly identify a poisonous plant at the scene of the poisoning accident.Methods Nineteen samples of leaves in 18 species of common poisonous plants of Ranunculaceae and Euphorbiaceaec were selected.Genomic DNA of these samples were extracted using improved hexadecyltrimethylammonium bromide (CTAB) method,species identification (genus level) were performed using random amplified polymorphic DNA (RAPD),intersimple sequences repeats (ISSR) and DNA barcoding,cluster analysis were performed using NYSTS,SPSS,PAUP,and MEGA software.Accuracy,reliability,timeliness,and operability were compared between the 3 molecular marker techniques.Results Accuracy:genus level of samples failed to be identified using RAPD ; and the accuracy rates in genus level of identification using ISSR and DNA barcoding were 68％ and 100％,respectively.Reliability:the subjective influence rates and the repetitive rates in RAPD and ISSR were 47％ and 26％ as well as 47％ and 45％,respectively; the success rates of amplification and sequencing of trnH-psbA fragment using DNA barcoding both were 100％.Timeliness:the time from extraction of DNA to accomplishment of identification in the 3 molecular marker techniques were 8.5,9.0,and 42.2 h,respectively.Operability:species identification using RAPD and ISSR could be done in a common laboratory; special sequencing equipment was needed when using DNA barcoding technique in the identification of poisonous plants.Conclusion The preliminary study results show that DNA barcoding is a molecular identification technique more suitable for rapid determination of the poisonous plant at the scene of the sudden poisoning accident.
