| AIDS患者合并感染耶氏肺孢子虫的 ITS1-5.8S rDNA-ITS2序列测定及基因型初步分析 |
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| 引用本文： | 林睿,黎学铭,张陆娟,秦英梅,欧阳颐,张鸿满,黄绍标,江河.AIDS患者合并感染耶氏肺孢子虫的 ITS1-5.8S rDNA-ITS2序列测定及基因型初步分析[J].广西医学,2008,30(12):1831-1833. |
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| 作者姓名： | 林睿 黎学铭 张陆娟 秦英梅 欧阳颐 张鸿满 黄绍标 江河 |
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| 作者单位： | 1. 广西壮族自治区疾病预防控制中心,南宁市,530028
2. 广西南宁市第四人民医院,南宁市,530023 |
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| 基金项目： | 广西科学研究与技术开发计划资金资助课题，广西医疗卫生重点科研课题 |
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| 摘 要： | 目的 测定AIDS患者痰液标本中耶氏肺孢子虫的ITS1-5.8S rDNA-ITS2序列并进行基因型分析.方法 收集AJDS患者痰液标本,二硫苏糖醇(DTT)消化处理后进行六亚甲基四胺银(GMS)染色镜检;提取肺孢子虫DNA,巢式PCR扩增后进行测序,用Blast程序和DNAMAN软件对所测序列进行同源性检索和序列间比对,并和SD大鼠卡氏肺孢子虫ITS1-5.8S rDNA-ITS2序列进行比较分析.结果 GMS染色法未检测到肺孢子虫,巢式PER成功扩增出4例(31%),其ITS1-5.8S rDNA-ITS2基因序列长度为469 bp,ITS1、5.8S rDNA、rTS2基因片段分别为146、158、165 bp,与SD大鼠ITS1-5.8S rDNA-ITS2的基因同源性为86%(152/176),其中多为5.8S rDNA同源86%(136/158),ITS基因间未发现显著性同源;病例间耶氏肺孢子虫ITS1-5.8S rDNA-ITS2的基因同源性为98%～99%,其中5.8S rDNA高度保守,ITS1基因存在4个差异位点并具规律性,据此把耶氏肺孢子虫初步分为A型和B型;ITS2基因存在3个基因位点,但差异无规律性.结论 成功从AIDS患者痰液标本中获取耶氏肺孢子虫ITS1-5.8S rDNA-ITS2基因序列,其和大鼠源基因差异明显,不同人源宿主耶氏肺孢子虫可存在不同基因型.
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| 关 键 词： | 痰液 耶氏肺孢子虫 内转录间隔区(ITS) 序列测定 基因型 |
Analysis of ITS1-5.8S rDNA-ITS2 sequence of Pneumocystis jirovecii isolated from AIDS patients |
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| LIN Rui,LI Xue-ming,ZHANG Lu-juan,QIN Ying-mei,OU Yang-yi,ZHANG Hong-man,HUANG Shao-biao,JIANG He.Analysis of ITS1-5.8S rDNA-ITS2 sequence of Pneumocystis jirovecii isolated from AIDS patients[J].Guangxi Medical Journal,2008,30(12):1831-1833. |
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| Authors: | LIN Rui LI Xue-ming ZHANG Lu-juan QIN Ying-mei OU Yang-yi ZHANG Hong-man HUANG Shao-biao JIANG He |
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| Institution: | LIN Rui1,LI Xue-ming1,ZHANG Lu-juan1,QIN Ying-mei2,OU Yang-yi1,ZHANG Hong-man1,HUANG Shao-biao2,JIANG He1(1 Guangxi Zhuang Autonomous Region Center for Disease Prevention , Control,Nanning 530028,China,2 The fourth People\'s Hospital of Nanning,Nanning 530023,China) |
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| Abstract: | Objective To examine the ITS1-5.8S rDNA-ITS2 sequence and analyze the genotype of Pneumocystis jirovecii(P.jirovecii) from the sputum specimens of AIDS patients.Methods Sputum specimens were collected from AIDS patients,then were digested with dithiothreitol(DTT),and stained with Gomori's methenamine silver stained(GMS).P.jirovecii were observed under microscope.The DNAs of P.jirovecii were extracted from sputum,the ITS1-5.8S rDNA-ITS2 of P.jirovecii were amplified by nested PCR.The PCR products were purifi... |
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| Keywords: | AJDs |
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