Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA ...Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA (SSUrRNA) of Endoreticulatus bombycis. The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned. It had a length of 1 230 nucleotides and a percentage GC content of 51.3%. The accession number in GenBank is gi11181769|AY009115. The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package. The secondary structure model revealed that Endoreticulatus bombycis lacked many eukaryotic helices, such as helix10, helix 11, helix 18, helix 43 and helix 46,but it has V4 region and has more of prokaryotic character. Analyzed by Blast, Endoreticulatus bombycis, Endoreticulatus schubergi and Pleistophora sp.(Sd-Nu-IW8201) have a high similarity, over 98%. Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd-Nu-IW8201) was in a group and the other Pleistophora species were in another group .展开更多
目的应用巢式PCR扩增小亚单位核糖体核糖核酸(18S r RNA)基因确诊卵形疟原虫感染。方法采集2012-2013年山东省疟疾患者全血和滤纸血样,吉氏染色镜检观察血膜疟原虫形态并鉴定虫种。提取血样中DNA,根据疟原虫18S r RNA基因以属和种特异...目的应用巢式PCR扩增小亚单位核糖体核糖核酸(18S r RNA)基因确诊卵形疟原虫感染。方法采集2012-2013年山东省疟疾患者全血和滤纸血样,吉氏染色镜检观察血膜疟原虫形态并鉴定虫种。提取血样中DNA,根据疟原虫18S r RNA基因以属和种特异性引物进行巢式PCR扩增,筛查卵形疟阳性标本并进行测序验证。结果2012和2013年山东省共筛查到7例输入性卵形疟原虫感染病例。巢式PCR扩增7份血样均在800 bp处出现卵形疟原虫特异性单一条带,Blast比对表明产物序列与Gen Bank卵形疟参考序列一致。结论经巢式PCR方法确诊了2012和2013年山东省7例输入性卵形疟病例。展开更多
文摘Endoreticulatus bombycis is a new pathogenic microspordia isolated from the silkworm larvae. With polymerase chain reaction (PCR) method,we amplified a fragment of the core sequence of the small subunit ribosomal RNA (SSUrRNA) of Endoreticulatus bombycis. The SSUrRNA fragment was inserted into pMD18-T Vector and then cloned. It had a length of 1 230 nucleotides and a percentage GC content of 51.3%. The accession number in GenBank is gi11181769|AY009115. The secondary structure model of the SSUrRNA of Endoreticulatus bombycis was constructed both on the bases of the sequence alignment and RNAFOLD program of the PC-GENE package. The secondary structure model revealed that Endoreticulatus bombycis lacked many eukaryotic helices, such as helix10, helix 11, helix 18, helix 43 and helix 46,but it has V4 region and has more of prokaryotic character. Analyzed by Blast, Endoreticulatus bombycis, Endoreticulatus schubergi and Pleistophora sp.(Sd-Nu-IW8201) have a high similarity, over 98%. Phylogeny tree of Endoreticulatus and Pleistophora species showed that Endoreticulatus and Pleistophora sp.(Sd-Nu-IW8201) was in a group and the other Pleistophora species were in another group .
文摘目的应用巢式PCR扩增小亚单位核糖体核糖核酸(18S r RNA)基因确诊卵形疟原虫感染。方法采集2012-2013年山东省疟疾患者全血和滤纸血样,吉氏染色镜检观察血膜疟原虫形态并鉴定虫种。提取血样中DNA,根据疟原虫18S r RNA基因以属和种特异性引物进行巢式PCR扩增,筛查卵形疟阳性标本并进行测序验证。结果2012和2013年山东省共筛查到7例输入性卵形疟原虫感染病例。巢式PCR扩增7份血样均在800 bp处出现卵形疟原虫特异性单一条带,Blast比对表明产物序列与Gen Bank卵形疟参考序列一致。结论经巢式PCR方法确诊了2012和2013年山东省7例输入性卵形疟病例。
