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
2013年
3期
140-143
,共4页
申继清%欧阳颐%王静%王鸽%黎学铭%陈乾容
申繼清%歐暘頤%王靜%王鴿%黎學銘%陳乾容
신계청%구양이%왕정%왕합%려학명%진건용
广州管圆线虫%内转录间隔区2%系统发生树%亲缘关系
廣州管圓線蟲%內轉錄間隔區2%繫統髮生樹%親緣關繫
엄주관원선충%내전록간격구2%계통발생수%친연관계
Angiostrongylus cantonensis%Internal transcribed spacer 2%Phylogenetic tree%Phylogenetic relationship
目的 分析和判定广西钦州、靖西、藤县三地广州管圆线虫(A ngiostrongylus cantonensis)分离株的基因差异和系统发生关系. 方法 从广西钦州、靖西、藤县采集野生蛞蝓,分离广州管圆线虫三期幼虫.提取广州管圆线虫三期幼虫的基因组DNA,特异性扩增内转录间隔区2(internal transcribed spacer 2,ITS2)基因并测序,与GenBank中的广州管圆线虫序列进行比对,分析基因变异.应用Mega5.05软件构建系统发生树,分析系统发生关系. 结果 从三地共采集623条蛞蝓,其中阳性蛞蝓共283条,平均感染率为45.4％.其中靖西感染率最高,为52.4％;藤县和钦州感染率分别为49.1％和19.3％.特异性扩增的ITS2长度为700 bp.GenBank检索获广州管圆线虫(A.cantone ns is)、A.dujardini以及脉居管圆线虫(A.vasorum)的相应序列为对照序列,枯病菌(Rhizoctonia solani)的相应序列为构建系统发生树的根(root)序列.广州管圆线虫钦州分离株、靖西分离株、藤县分离株与GenBank中的广州管圆线虫序列的同源性分别为99.5％、99.5％和99.7％;靖西分离株与藤县分离株、GenBank中广州管圆线虫序列的同源性分别为100％和99.5％;藤县分离株与及GenBank中广州管圆线虫序列的同源性为99.5％.三个分离株与A.dujardini、脉居管圆线虫(A.vasorum)的同源性低.以广州管圆线虫为标准株,靖西分离株有11处碱基变异,变异率为1.93％;钦州分离株有6处,变异率为1.05％;藤县分离株有15处,变异率为2.63％,TG短序列重复插入突变明显.在最大似然法(maximum likelihood method)和邻接法(neighbor-joining method)构建的系统发生树上,靖西、钦州及藤县的分离株与广州管圆线虫同属一个分枝,系统发生关系较近,A.dujardini、脉居管圆线虫居于另一分枝. 结论 广西三地的广州管圆线虫分离株存在较高的遗传多态性,与GenBank中的广州管圆线虫同源性高,亲缘关系近,初步判断三个分离株与GenBank中广州管圆线虫为同一虫种.
目的 分析和判定廣西欽州、靖西、籐縣三地廣州管圓線蟲(A ngiostrongylus cantonensis)分離株的基因差異和繫統髮生關繫. 方法 從廣西欽州、靖西、籐縣採集野生蛞蝓,分離廣州管圓線蟲三期幼蟲.提取廣州管圓線蟲三期幼蟲的基因組DNA,特異性擴增內轉錄間隔區2(internal transcribed spacer 2,ITS2)基因併測序,與GenBank中的廣州管圓線蟲序列進行比對,分析基因變異.應用Mega5.05軟件構建繫統髮生樹,分析繫統髮生關繫. 結果 從三地共採集623條蛞蝓,其中暘性蛞蝓共283條,平均感染率為45.4％.其中靖西感染率最高,為52.4％;籐縣和欽州感染率分彆為49.1％和19.3％.特異性擴增的ITS2長度為700 bp.GenBank檢索穫廣州管圓線蟲(A.cantone ns is)、A.dujardini以及脈居管圓線蟲(A.vasorum)的相應序列為對照序列,枯病菌(Rhizoctonia solani)的相應序列為構建繫統髮生樹的根(root)序列.廣州管圓線蟲欽州分離株、靖西分離株、籐縣分離株與GenBank中的廣州管圓線蟲序列的同源性分彆為99.5％、99.5％和99.7％;靖西分離株與籐縣分離株、GenBank中廣州管圓線蟲序列的同源性分彆為100％和99.5％;籐縣分離株與及GenBank中廣州管圓線蟲序列的同源性為99.5％.三箇分離株與A.dujardini、脈居管圓線蟲(A.vasorum)的同源性低.以廣州管圓線蟲為標準株,靖西分離株有11處堿基變異,變異率為1.93％;欽州分離株有6處,變異率為1.05％;籐縣分離株有15處,變異率為2.63％,TG短序列重複插入突變明顯.在最大似然法(maximum likelihood method)和鄰接法(neighbor-joining method)構建的繫統髮生樹上,靖西、欽州及籐縣的分離株與廣州管圓線蟲同屬一箇分枝,繫統髮生關繫較近,A.dujardini、脈居管圓線蟲居于另一分枝. 結論 廣西三地的廣州管圓線蟲分離株存在較高的遺傳多態性,與GenBank中的廣州管圓線蟲同源性高,親緣關繫近,初步判斷三箇分離株與GenBank中廣州管圓線蟲為同一蟲種.
목적 분석화판정엄서흠주、정서、등현삼지엄주관원선충(A ngiostrongylus cantonensis)분리주적기인차이화계통발생관계. 방법 종엄서흠주、정서、등현채집야생활유,분리엄주관원선충삼기유충.제취엄주관원선충삼기유충적기인조DNA,특이성확증내전록간격구2(internal transcribed spacer 2,ITS2)기인병측서,여GenBank중적엄주관원선충서렬진행비대,분석기인변이.응용Mega5.05연건구건계통발생수,분석계통발생관계. 결과 종삼지공채집623조활유,기중양성활유공283조,평균감염솔위45.4％.기중정서감염솔최고,위52.4％;등현화흠주감염솔분별위49.1％화19.3％.특이성확증적ITS2장도위700 bp.GenBank검색획엄주관원선충(A.cantone ns is)、A.dujardini이급맥거관원선충(A.vasorum)적상응서렬위대조서렬,고병균(Rhizoctonia solani)적상응서렬위구건계통발생수적근(root)서렬.엄주관원선충흠주분리주、정서분리주、등현분리주여GenBank중적엄주관원선충서렬적동원성분별위99.5％、99.5％화99.7％;정서분리주여등현분리주、GenBank중엄주관원선충서렬적동원성분별위100％화99.5％;등현분리주여급GenBank중엄주관원선충서렬적동원성위99.5％.삼개분리주여A.dujardini、맥거관원선충(A.vasorum)적동원성저.이엄주관원선충위표준주,정서분리주유11처감기변이,변이솔위1.93％;흠주분리주유6처,변이솔위1.05％;등현분리주유15처,변이솔위2.63％,TG단서렬중복삽입돌변명현.재최대사연법(maximum likelihood method)화린접법(neighbor-joining method)구건적계통발생수상,정서、흠주급등현적분리주여엄주관원선충동속일개분지,계통발생관계교근,A.dujardini、맥거관원선충거우령일분지. 결론 엄서삼지적엄주관원선충분리주존재교고적유전다태성,여GenBank중적엄주관원선충동원성고,친연관계근,초보판단삼개분리주여GenBank중엄주관원선충위동일충충.
Objective To analyze the phylogenetic relationship ofA ngiostrongylus cantonensts isolated from Qinzhou,Jingxi and Tengxian counties of Guangxi.Methods Slugs were collected from Qinzhou,Jingxi and Tengxian counties of Guangxi and the third stage larvae ofA ngiostrongylus cantonensis were collected from positive samples The genomic DNA was extracted from A ngiostrongylus cantonensis.Internal transcribed spacer 2 (ITS2) was amplified and the amplicon was sequenced,then analysis was carried out and compared with those from GenBank.Phylogenetic trees were constructed and phylogenetic relationship ofA ngiostrongylus cantonensis isolates was analyzed by Mega5.05 software.Results Totally 623 slugs from 3 counties were collected and 283 were infected with Angiostrongylus cantoncnsis with average infection rate of 45.4％.Among three counties,the infection rate in Jingxi was the highest with 52.4％,followed by Tengxian and Qinzhou with 49.1％ and 19.3％,respectively.Fragments of 700bp were specifically amplified.Corresponding sequence of A.cantonensis,A.dujardini and A.vasorum from GenBank were used as similarity compared sequence while Rhizoctonia solani as root sequence.Qinzhou.Jingxi and Tengxian isolates was very like to A.cantonensis from GenBank with homology of 99.5％,99.5％ and 99.7％,respectively.The homology between Jingxi,Tengxian isolates and A.cantonensis was 100％ and 99.5％,respectively,and 99.5％ between Tengxian isolates and A.cantonensis.The homology between the three isolates and A.dujardini,A.vasorumwere was low.Compared with A.cantonensis from GenBank,11 bp heteromorphosis was found in Jingxi isolate with the ratio of 1.93％,6bp in Qinzhou isolate with the ratio of 1.05％ and 15 bp in Tengxian isolate with the ratio of 2.63％.Reduplicated TG acids were obvious.In the phylogenetic trees constructed by maximum likelihood method and neighbor-joining method,Jingxi,Qinzhou,Tengxian isolates and A.cantonensis from GenBank were separated into one branch with closer phenogenetic relationship,while A.dujardini and A.vasorum were separated into another branch with far phenogenetic relationship from the three isolates.Conclusion The three isolates from Guangxi were similar to A.cantonensis from GenBank with higher homology,closer phenogenetic relationship and genetic variation.
