江西省HIV阳性人群隐孢子虫感染现状及基因特征分析  

Prevalence and genetic characteristics of Cryptosporidium infections among HIV⁃positive individuals in Jiangxi Province

作　　者：胡主花[1] 路亮[1] 俞英昉 李琳 王伟[1] 樊国印[1] 冯长华[1] 郑扬云[1] 彭国华[1] HU Zhuhua;LU Liang;YU Yingfang;LI Lin;WANG Wei;FAN Guoyin;FENG Changhua;ZHENG Yangyun;PENG Guohua(Nanchang Municipal Center for Disease Control and Prevention,Base of National Key Laboratory for the Prevention and Control of Infectious Diseases,Nanchang,Jiangxi 330038,China;National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research),National Health Commission Key Laboratory of Parasite and Vector Biology,WHO Collaborating Centre for Tropical Diseases,National Center for International Research on Tropical Diseases,Ministry of Science and Technology,China)

机构地区：[1]江西省南昌市疾病预防控制中心、传染病预防控制国家重点实验室研究基地,江西南昌330038 [2]中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心)、国家卫生健康委员会寄生虫病原与媒介生物学重点实验室、WHO热带病合作中心、科技部国家级热带病国际联合研究中心

出　　处：《中国血吸虫病防治杂志》2024年第6期637-642,共6页Chinese Journal of Schistosomiasis Control

基　　金：江西省南昌市科技支撑重点项目(洪科字[2021]129号-20);江西省卫生健康委员会科技计划(202140184);江西省卫生健康委员会科技计划(202311341)。

摘　　要：目的了解江西省HIV阳性人群隐孢子虫感染现状及虫种和基因型分布。方法2022年1月—2023年3月,于江西省3家艾滋病治疗定点医院收集HIV阳性人群社会人口学特征和临床资料,并采集其新鲜粪便样本。提取粪样基因组DNA,采用基于隐孢子虫小亚基核糖体RNA(small subunit ribosomal RNA,SSU r RNA)基因的巢式PCR方法检测样本,阳性样本再扩增其隐孢子虫60 k Da糖蛋白(60 k Da glycoprotein,gp60)基因。阳性样本两种基因均重复进行3次PCR扩增。采用1.5%琼脂糖凝胶电泳检测第二轮PCR扩增产物,对两基因的疑似阳性产物进行双向基因测序和比对。应用MEGA 11.0软件基于邻接法绘制系统进化树,以鉴定隐孢子虫虫种、基因型和基因亚型。结果累计将382例江西省HIV阳性者纳入研究,其中2例感染隐孢子虫,感染率为0.52%(2/382),感染者均无腹痛、腹泻等临床症状。经测序与序列比对,2例感染样本隐孢子虫分离株与火鸡隐孢子虫分离株相似性分别为99.76%和99.88%。基于隐孢子虫SSU rRNA基因序列的系统进化树分析显示,2份隐孢子虫阳性样本虫种均为火鸡隐孢子虫。对2株火鸡隐孢子虫gp60基因进行巢式PCR扩增、测序和序列比对,分别鉴定为火鸡隐孢子虫Ⅲe A17G2R1亚型和Ⅲb A25G1R1a亚型,其中Ⅲb A25G1R1a亚型为人体中首次发现。结论江西省HIV阳性人群隐孢子虫感染率较低。临床治疗中应关注无腹泻症状的HIV阳性者隐孢子虫感染可能性。Objective To investigate the prevalence of Cryptosporidium infection and the distribution of parasite species and genotypes among HIV⁃positive individuals in Jiangxi Province.Methods HIV⁃positive individuals'sociodemographic and clinical data were collected from three AIDS designated hospitals in Jiangxi Province from January 2022 to March 2023.Subjects'stool samples were collected,and genomic DNA was extracted from stool samples.Nested PCR assay was performed based on the small subunit ribosomal RNA(SSU rRNA)gene of Cryptosporidium,and Cryptosporidium gp60 gene was amplified in stool samples positive for the SSU rRNA gene.The second⁃round PCR amplification product was checked with 1.5%agarose gel electrophoresis,and the products of suspected positive amplifications were sequenced,followed by sequence alignment.The phylogenetic tree was created using the Neighbor⁃Joining method with the software MEGA 11.0,to characterize the species,genotypes and sub⁃genotypes of Cryptosporidium.Results A total of 382 HIV⁃positive individuals were enrolled,with two cases identified with Cryptosporidium infection(0.52%prevalence),and both cases had no abdominal pain or diarrhea.Following sequencing and sequence alignment,the gene sequences of these two Cryptosporidium isolates shared 99.76%and 99.88%similarity with the gene sequence of C.meleagridis isolates.Phylogenetic analysis based on the Cryptosporidium SSU rRNA gene sequence identified the species of these two Cryptosporidium⁃positive stool samples as C.meleagridis.Following nested PCR amplification of the Cryptosporidium gp60 gene,sequencing and sequence alignment,the two C.meleagridis isolates were characterized as ⅢeA17G2R1 and ⅢbA25G1R1a sub⁃genotypes,and the sub⁃genotype ⅢbA25G1R1a was firstly described in humans.Conclusion The prevalence of Cryptosporidium is low among HIV⁃positive individuals in Jiangxi Province.The likelihood of Cryptosporidium infection cannot be neglected among HIV⁃positive individuals without diarrhea.

关 键 词：隐孢子虫 HIV 基因特征 江西省 

分 类 号：R382.3[医药卫生—医学寄生虫学]