收稿日期: 2021-08-20
修回日期: 2021-09-16
网络出版日期: 2022-03-31
基金资助
国家科技重大专项(2018ZX10101002);WHO示范项目(UNOPS/AND/G/2016/01);河南省科技攻关项目(182102310671)
Current prevalence and sub-genotype analysis of Blastocystis in residents of Luoyang City
Received date: 2021-08-20
Revised date: 2021-09-16
Online published: 2022-03-31
Supported by
National Science and Technology Major Project(2018ZX10101002);WHO Demo Project(UNOPS/AND/G/2016/01);Henan Province Science and Technology Research Project(182102310671)
目的 了解洛阳市人群芽囊原虫感染情况及基因亚型分布。 方法 2020年9月13—20日,采取整群随机抽样的方法选择洛阳市土源性线虫病流动监测点中的洛宁县作为调查点,分别在程家庄、西街村、中高村、南坡村、磨沟村等5个行政村抽取3周岁以上常住居民不少于200人。采集调查对象基本信息,包括姓名、性别、年龄、民族、职业、文化程度、村庄和调查前3个月内是否服用驱虫药。调查芽囊原虫感染者的生活卫生条件及临床体征等信息。提取感染者粪样DNA,PCR扩增芽囊原虫小亚基RNA（SSU rRNA）基因,测序后在GenBank数据库中进行BLAST比对,鉴定扩增序列所属基因亚型。采用Mega 7.0最大似然法构建基于SSU rRNA基因的系统发育树。采用SPSS 25.0统计学软件进行统计分析。 结果 共调查890人,检出芽囊原虫阳性20例,总感染率为2.2%（20/890）。其中男性感染率为2.0%（8/399）,女性感染率为2.4%（12/491）,二者差异无统计学意义（χ2 = 0.193,P > 0.05）。11~20岁年龄组感染率最高,为5.1%（3/59）,不同年龄组间感染率差异有统计学意义（Fisher = 11.895,P < 0.05）。农民感染率最高,为2.6%（17/653）,不同职业感染率差异无统计学意义（Fisher = 1.535,P > 0.05）。初中文化程度感染率最高,为 2.7%（12/452）,不同文化程度感染率差异无统计学意义(Fisher = 1.283,P > 0.05）。中高村居民芽囊原虫感染率最高,为4.3%(7/164),5个行政村居民芽囊原虫感染率差异有统计学意义（Fisher = 10.929,P < 0.05）。PCR扩增结果显示,20份样品扩增出芽囊原虫SSU rRNA基因片段,大小约1 100 bp。测序结果以ST3基因亚型为主,占90.0%（18/20）,ST1基因型占10.0%（2/20）。磨沟村和中高村的感染者包括ST3和ST1基因亚型,南坡村和西街村的感染者均为ST3基因亚型。系统进化树结果显示,20份阳性粪样中,有18份与奥地利分离株（GenBank登录号MN914073.1）、德国分离株（GenBank登录号MK801403.1）和新加坡分离株（GenBank登录号KX618192.1）处于同一个进化分支,均为ST3基因亚型;2份分别与德国分离株（GenBank登录号MK801408.1）和奥地利分离株（GenBank登录号MN914072.1）处于同一个分支,为ST1基因亚型,测序结果一致。对20例感染者进行卫生条件及临床体征调查结果显示,使用旱厕或粪池者占65.0%（13/20）,使用水冲厕所者占35.0%（7/20）;家中饲养宠物或家畜者占45.0%（9/20）;3例有腹痛、腹泻、腹胀等症状,分别为1例ST1亚型、2例ST3亚型;4例有大便干结等症状,分别为1例ST1基因亚型,3例ST3基因亚型。 结论 洛阳市人群芽囊原虫总感染率较低,主要为ST3 基因亚型。
陈慧慧 , 李云霞 , 宁超群 , 邓艳 , 张红卫 , 韦雪芳 , 锁慧芳 , 张颋 , 刘琴 , 陈军虎 , 田利光 . 洛阳市人群芽囊原虫感染现状及基因亚型分析[J]. 中国寄生虫学与寄生虫病杂志, 2022 , 40(2) : 216 -222 . DOI: 10.12140/j.issn.1000-7423.2022.02.013
Objective To understand the infection status and distribution of sub-genotype of Blastocystis hominis in Luoyang City, Henan Province. Methods From September 13-20, 2020, a cross-sectional study with cluster random sampling was conducted in Luoning County, which was selected from the Luoyang City soil-transmitted nematode surveillance sites as the survey site. In the study, we selected no less than 200 permanent residents aged 3 and above from five administrative villages of Cheng jia zhuang, Xijie, Zhong gao, Nanpo and Mogou. A questionnaire was performed to collect the basic information of name, gender, age, ethnicity, occupation, education level, village and whether they had taken deworming medication in the three months prior to the survey. For those infected with Blastocystis information on living hygiene conditions and clinical signs were investigated. Faecal DNA was extracted and the small subunit RNA (SSU-rRNA) gene was amplified by PCR, sequenced and BLAST in the GenBank database to identify the genetic subtype. A phylogenetic tree based on the SSU-rRNA gene was constructed using the Mega 7.0, and allstatistical analysis was performed using SPSS 25.0 software; Results A total of 890 people were investigated and 20 people were found positive for Blastocystis infection. The Blastocystis infection rate was 2.2% (20/890). Among them, the male infection rate was 2.0% (8/399), and the female infection rate was 2.4% (12/491), and the difference was not statistically significant(χ2 = 0.193, P > 0.05). The 11-20 years age group had the highest infection rate, which was 5.1% (3/59), and the difference in infection rate among different age groups was statistically significant(Fisher =11.895, P < 0.05). Farmers had the highest infection rate at 2.6%, and there was no significant difference in infection rates among different occupations(Fisher = 1.535, P > 0.05). The infection rate among junior high school educated participants was the highest (2.7%), and there was no significant difference in infection rate among different education levels(Fisher = 1.283, P > 0.05). We found the Blastocystis infection rate in Zhonggao Village was the highest at 4.3% (7/164), and differences among 5 administrative villages were statistically significant (Fisher = 10.929, P < 0.05). As for PCR amplification, results showed that 20 samples amplified the SSU-rRNA gene fragment of Blastocystis, about 1 100 bp in size. Sequencing results showed that ST3 subtypes accounted for 90.0% (18/20), and ST1 subtypes accounted for 10.0% (2/20), which was consistent with the phylogenetic tree results. Within the infectious of Blastocystis, Mogou and Zhonggao villages included both ST3 subtypes and ST1 subtypes, and Nanpo and Xijie villages were both ST3 subtypes. Then, the phylogenetic tree results showed that 18 of the 20 positive faecal samples were in the same evolutionary branch as the Austrian isolate (GenBank accession no. MN914073.1), the German isolate (GenBank accession no. MK801403.1) and the Singaporean isolate (GenBank accession no. KX618192.1), all of which were ST3 subtypes. Two copies were in the same evolutionary branch as the German isolate (GenBank accession no. MK801408.1) and the Austrian isolate (GenBank accession no. MN914072.1), both ST1 subtypes of Blastocystis. As for hygienic conditions and clinical manifestations in 20 infected patients, 65.0% (13/20) used dry toilets or cesspools, 35.0% (7/20) used flushing toilets, and 45.0% (9/20) kept pets or livestock at home. 15.0% (3/20) showed symptoms of abdominal pain, diarrhea and abdominal distension, including one ST1 subtype and two ST3 subtypes; 20% (3/20) showed symptoms of dry stools, including one ST1 subtype and three ST3 subtypes. Conclusion The overall infection rate of B. hominis in the population of Luoyang City is relatively low, with the main sub-genotype ST3.
Key words： Blastocystis sp.; Subtype; Epidemiological investigation; Luoyang City
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