中国寄生虫学与寄生虫病杂志 ›› 2013, Vol. 31 ›› Issue (5): 19-410-411.

• 研究简报 • 上一篇    下一篇

云南HIV阳性患者感染弓形虫基因型的初步分析

聂大平1,2,贾玉玺1,陈凌娟1,尤英霞1,李伟1,申丽洁3 *   

  1. 1 大理学院基础医学院寄生虫学教研室,大理671003;2 江西省吉安县人民医院,吉安343100;
    3 昆明医科大学病原生物学与免疫学系,昆明650500
  • 出版日期:2013-10-30 发布日期:2014-07-24

Genotypes of Toxoplasma gondii Isolates from HIV Positive Patients in Yunnan Province

NIE Da-ping1,2,JIA Yu-xi1,CHEN Ling-juan1,YOU Ying-xia1,LI Wei1,SHEN Li-jie3 *   

  1. 1 Department of Parasitology,College of Basic Medicine,Dali University,Dali 671003,China;2 Ji’an County People’s Hospital of Jiangxi Province,Ji’an 343100,China;3 Department of Pathogen Biology and Immunology,Kunming Medical University,Kunming 650500,China
  • Online:2013-10-30 Published:2014-07-24

摘要: 2011年9月至2012年12月收集云南省西部地区HIV阳性患者全血样品150份,利用巢氏PCR技术对血样进行弓形虫B1基因的扩增,扩增产物经限制性内切酶Pm1Ⅰ和XhoⅠ酶切,进行限制性内切酶片段长度多态性分析(RFLP)。自B1基因扩增阳性的血样中取5份扩增产物测序,结果与标准基因Ⅰ型株B1基因序列(登录号为AF179871)进行比对分析。在150份HIV阳性患者血样中,13份扩增出弓形虫B1基因,约530 bp。PCR产物酶切结果显示,13份血样均得到1个片段,约为500 bp,与标准基因Ⅰ型株的条带一致。测序结果显示,4份血样的扩增产物完全一致,与基因Ⅰ型株的B1基因序列仅有3个碱基的差异;余1份血样的扩增产物在第230对碱基处G→A。因此初步确定此次研究中的云南省西部地区HIV阳性患者感染弓形虫基因型为Ⅰ型。

关键词: 人类免疫缺陷病毒, 刚地弓形虫, B1基因, 巢式PCR, 基因分型

Abstract:  One hundred and fifty serum samples of HIV positive patients were collected in western Yunnan Province from September 2011 to December 2012. Toxoplasma gondii B1 gene was amplified by nested PCR. Genotyping of T. gondii isolates were performed by restriction fragment length polymorphism(RFLP) with Pm1Ⅰ and XhoⅠ. 13 samples were found positive with the B1 gene (530 bp) amplification and belonged to type Ⅰ. The sequencing results showed that 4 T. gondii B1 gene positive samples were identical, with 3 nucleotide variation compared with T. gondii strain RH (type Ⅰ) B1 gene (GenBank No. AF179871), and in the other sample a “G→A” mutation at 230 bp was detected. The results indicated that the genotype of Toxoplasma gondii in HIV positive patients in Yunnan Province is type Ⅰ.

Key words: HIV, Toxoplasma gondii, B1 gene, Nested PCR, Genotyping