黔东南地区侗族人群芽囊原虫感染情况及影响因素分析

doi:10.12140/j.issn.1000-7423.2023.06.009

中国寄生虫学与寄生虫病杂志 ›› 2023, Vol. 41 ›› Issue (6): 725-732.doi: 10.12140/j.issn.1000-7423.2023.06.009

黔东南地区侗族人群芽囊原虫感染情况及影响因素分析

李琳¹(), 朱爱娅², 蔡玉春¹, 俞铖航¹, 陈木新¹, 田利光¹^,^*()

1 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）；传染病溯源预警与智能决策全国重点实验室；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；世界卫生组织热带病合作中心；科技部国家级热带病国际研究中心，上海 200025
2 贵州省疾病预防控制中心，贵阳 550004

收稿日期:2023-07-11 修回日期:2023-09-27 出版日期:2023-12-30 发布日期:2023-12-22
通讯作者: * 田利光（1970-），男，博士，研究员，从事寄生虫病流行病学研究。E-mail：jztlg@126.com
作者简介:李琳（1998-），女，硕士研究生。从事寄生虫病流行病学研究。E-mail：li13789360978@163.com
基金资助:
上海市自然科学基金(21ZR1469900);国家科技基础条件平台国家寄生虫资源库项目(NPRC-2019-194-30);贵州省科技计划项目（黔科合支撑〔2023〕一般183)

Prevalence and influence factors of Blastocystis infection in residents of Dong ethnic group in Southeast Guizhou Province

LI Lin¹(), ZHU Aiya², CAI Yuchun¹, YU Chenghang¹, CHEN Muxin¹, TIAN Liguang¹^,^*()

1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; Key Laboratory on Parasite and Vector Biology, Ministry of Health; WHO Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai 200025, China
2 Center for Disease Control and Prevention of Guizhou Province, Guiyang 550004, China

Received:2023-07-11 Revised:2023-09-27 Online:2023-12-30 Published:2023-12-22
Contact: * E-mail: jztlg@126.com
Supported by:
Natural Science Foundation of Shanghai(21ZR1469900);National Parasitic Resources Center, and the Ministry of Science and Technology fund(NPRC-2019-194-30);Guizhou Provincial Scientific and Technological Program [(Qian Ke He (2023) General]

摘要/Abstract

摘要：

目的了解2022年贵州省侗族人群芽囊原虫感染情况及影响因素。方法于2022年采用横断面调查方法，对黔东南侗族地区黎平县芒岭村和地青村居民进行问卷调查，包括一般人口学信息和卫生习惯、有无胃肠道症状等，并采集粪样，提取粪样芽囊原虫基因组DNA，采用PCR扩增芽囊原虫小亚基核糖体RNA（SSU rRNA）基因片段序列。取阳性产物测序，测序结果在GenBank数据库中进行BLAST序列比对，确定芽囊原虫感染基因亚型。采用MEGA 7.0软件，用邻接法构建基于芽囊原虫SSU rRNA基因的系统进化树。采用卡方检验和Fisher确切概率法对纳入本研究居民芽囊原虫感染影响因素进行统计学分析。结果共调查了329名居民，女性200人（60.8%）、男性129人（39.2%），平均年龄（47.95 ± 20.39）岁，侗族占98.5%（324/329）、汉族占1.2%（4/329）。PCR结果显示，有16份粪样扩增出芽囊原虫SSU rRNA基因片段，大小约1 100 bp。芽囊原虫总阳性率为4.9%（16/329）。其中男性阳性率为3.1%（4/129），女性阳性率为6.0%（12/200），二者差异无统计学意义（χ² = 1.425，P > 0.05）。> 69岁年龄组阳性率较高，为8.1%（3/37），不同年龄组间阳性率差异无统计学意义（χ² = 3.927，P > 0.05）。芽囊原虫阳性者职业均为农民，阳性率为6.1%（16/261），不同职业阳性率差异无统计学意义（χ² = 3.865，P > 0.05）。家庭人口数为6人及以上居民阳性率最高，为12.7%（7/55），不同家庭人口数阳性率差异有统计学意义（Fisher = 13.957，P < 0.01）。测序结果显示，芒岭村和地青村居民感染的芽囊原虫基因亚型有ST2、ST3、ST6、ST10等4种，其中ST6型最多（8/16），其次为ST3型（4/16）。BLAST比对分析结果显示，16份芽囊原虫阳性序列与参照序列的序列相似性均为90%以上。系统进化树结果显示，各基因亚型与其参照序列聚为一支，亲缘性较近。单因素分析结果显示，饮用水来源、家庭人口数、每日排便次数和大便性状对芽囊原虫感染的差异有统计学意义（χ² = 8.778、9.552、7.384、15.680，均P < 0.05）。多因素分析结果表明，饮用非开水[OR = 18.347，95% CI：（3.754，89.673）]是农村居民感染芽囊原虫的危险因素。结论贵州省黔东南地区侗族居民的芽囊原虫总阳性率较高，主要基因亚型为ST6。饮用非开水是农村居民感染芽囊原虫的主要危险因素。

关键词: 芽囊原虫, 亚型, 感染率, 影响因素, 少数民族

Abstract:

Objective To investigate the prevalence and influence factors of Blastocystis in the Dong ethnic group in Guizhou Province in 2022. Methods A cross-sectional survey was conducted through questionnaire investigation in the residents of Mangling Village and Diqing Village in Liping County, Dong ethnic group region, southeast Guizhou Province in 2022. The survey collected general demographic information, hygienic habits, evidence of gastrointestinal symtoms. In addition, fecal samples were collected to extract Blastocystis DNA from which the SSU rRNA gene fragment was amplified by PCR. The positive ampicon was sequenced, and the results were aligned by BLAST in the GenBank database to determine the subgenotype of Blastocystis. The MEGA7.0 software was used to construct phylogenetic tree based on the Blastocystis SSU rRNA gene sequence by neighbor-joining method. Chi-square test and Fisher exact probability method were used to analyze the influencing factors of Blastocystis infection in the residents included in this study. Results A total of 329 village residents were surveyed, including 200 females (60.8%) and 129 males (39.2%), with an average age of 47.95 ± 20.39 years, of which 98.5% (324/329) were of Dong ethnic group, and 1.2% (4/329) were of Han ethnic group. PCR results showed that 16 fecal samples amplified Blastocystis SSU rRNA fragments with a size of about 1 100 bp. The overall Blastocystis infection rate was 4.9% (16/329). There was no significant difference in the infection rates in males (3.1%, 3/129) and females (6.0%, 12/200) (χ² = 1.425, P > 0.05). The infection rate was higher in the age group over 69 years old, 8.1% (3/37), and there was no significant difference in the infection rate among different age groups (χ² = 3.865, P > 0.05). The Blastocystis infected individuals were all farmers, and the infection rate was 6.1% (16/261), and there was no significant difference in the infection rate between different occupations (χ² = 3.157, P > 0.05). The infection rate was the highest in households with 6 or more people (12.7%, 7/55), and the difference in infection rate between different household sizes was statistically significant (Fisher = 13.957, P < 0.01). There were 4 genotypes (ST2, ST3, ST6, ST10) infecting the residents of Mangling Village and Diqing Village. Among them, sub-genotype 6 is the most common, accounting for 50.0% (8/16), followed by sub-genotype 3, accounting for 25.0% (4/16). BLAST comparison analysis showed that the similarity between the positive sequences of 16 Blastocystis and the reference sequences was more than 90%. The phylogenetic tree results also showed that each gene subtype clustered with its reference sequence, and the parentage was high. Univariate analysis showed that drinking water source, family size, daily stool frequency and stool traits have significant differences in Blastocystis infection (χ² = 8.778, 9.552, 7.384, 15.680; P < 0.05). The results of multivariate analysis showed that drinking unboiled water [OR = 18.347, 95% CI: (3.754, 89.673)] was a risk factor for Blastocystis infection in rural residents. Conclusion The overall infection rate of Blastocystis among Dong ethnic residents in southeast Guizhou Province is comparatively, with the main subgenotype ST6. Drinking unboiled water is the main risk factor for infection with Blastocystis in rural residents.

Key words: Blastocystis sp., Subtype, Infection rate, Influence factor, Minority

中图分类号:

R531.9

李琳, 朱爱娅, 蔡玉春, 俞铖航, 陈木新, 田利光. 黔东南地区侗族人群芽囊原虫感染情况及影响因素分析[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(6): 725-732.

LI Lin, ZHU Aiya, CAI Yuchun, YU Chenghang, CHEN Muxin, TIAN Liguang. Prevalence and influence factors of Blastocystis infection in residents of Dong ethnic group in Southeast Guizhou Province[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2023, 41(6): 725-732.

图/表 3

表1

黔东南地区侗族人群芽囊原虫感染影响因素单因素分析

影响因素 Variable	调查人数 No. Investigate	感染人数 No. infected	感染率/% Infection rate/%	χ²值 χ² value	P 值 P value
性别 Gender				1.425	> 0.05
男 Male	129	4	3.1
女 Female	200	12	6.0
年龄/岁 Age/Year				3.865	> 0.05
≤ 17	49	0	0
18~45	58	4	6.9
46~69	185	9	4.9
> 69	37	3	8.1
职业 Occupation				3.157	> 0.05
农民 Farmer	261	16	6.1
非农民 Non-farmer	68	0	0
受教育程度 Educational level				4.256	> 0.05
文盲 Illiteracy	52	3	5.8
小学 Primary school	202	7	3.5
初中及以上 Junior high school and above	75	6	8.0
家庭人口数 No. family				9.552	< 0.05
3人及以下 Three persons and below	52	2	3.9
4人 Four persons	136	3	2.2
5人 Five persons	86	4	4.7
6人及以上 Six persons and above	55	7	12.7
厕所类型 Toilet type				0.021^b	> 0.05
旱厕 Latrine	10	1	1/10
粪池 Stercorary	12	1	1/12
水冲厕 Flush toilet	307	14	4.6
饮用水来源 Source of drinking water				8.778	< 0.01
开水 Boiled water	180	3	1.7
非开水 Non-boiled water	149	13	8.7
是否饲养家畜/禽 Whether to keep livestock/poultry				0.008	> 0.05
是 Yes	275	12	4.4
否 No	54	4	7.4
接触家畜/禽的频率 Frequency of contact with livestock/poultry				3.883	> 0.05
每天接触 Daily	191	9	4.7
每周接触3~4次 Three-four times a week	23	2	8.7
每周接触1~2次 One-two times a week	31	3	9.7
不接触 Never	84	2	2.4
用餐前洗手 Wash hands before eating				3.831	> 0.05
偶尔 Occasionally	86	3	3.5
经常 Frequently	177	9	5.1
每次 Every time	66	4	6.1
使用手清洁剂 Use cleaner				23.647^a	> 0.05
从不 Never	17	1	1/17
偶尔 Occasionally	244	12	4.9
经常 Frequently	36	3	0.8
每次 Every time	32	0	0
近3个月是否有临床症状 Whether there have been clinical symptoms in the last 3 months				3.563	> 0.05
是 Yes	42	2	4.8
否 No	287	14	4.9
是否有食欲不振、呕吐 Loss of appetite, vomiting				0	> 0.05
是 Yes	32	2	6.3
否 No	297	14	4.7
每日排大便次数 Number of bowel movements per day				7.384^a	< 0.05
1~2次 One-two times	312	14	4.5
3~4次 Three-four times	12	0	0
2~3天一次 Once every two-three days	5	2	2/5
大便性状 Character of stool				15.68	< 0.01
软便 Soft stool	264	8	3.0
干结便 Dry stool	41	3	7.3
稀便 Loose stool	24	5	20.8

表1

表2

图1

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变量 Variable	赋值 Variable assignment	回归系数Coefficient	标准误 Standard error	Wald χ²	P值 P value	OR（95% CI）
职业 Occupation	非农民 = 0，农民 = 1 Non-farmer = 0，Farmer = 1	18.033	4 335.06	0	> 0.05	—
饮用水来源 Source of drinking water	开水 = 0，非开水 = 1 Boiled water = 0，Non-boiled water = 1	2.909	0.81	12.916	< 0.01	18.347（3.754，89.673）
近3个月是否有临床症状 Whether there have been clinical symptoms in the last 3 months	否 = 0，是 = 1 No = 0，Yes = 1	-1.358	0.811	2.800	> 0.05	0.257（0.052，1.262）
家庭人口数 No. family	3人及以下 = 0 Three persons and below = 0			6.765	> 0.05
	4人 = 1 Four persons = 1	-1.252	1.014	1.524	> 0.05	0.286（0.039，2.088）
	5人 = 2 Five persons = 2	0.038	0.940	0.020	> 0.05	1.038（0.165，6.548）
	6人及以上 = 3 Six persons and above = 3	1.014	0.979	1.071	> 0.05	2.758（0.404，18.791）
大便性状 Character of stool	软便 = 0 Soft stool = 0			8.224	< 0.05
	稀便 = 1 Loose stool = 1	2.083	0.810	6.614	< 0.01	8.028（1.641，39.269）
	干结便 = 2 Dry stool = 2	1.585	0.756	4.394	< 0.05	4.880（1.109，21.480）

黔东南地区侗族人群芽囊原虫感染情况及影响因素分析

Prevalence and influence factors of Blastocystis infection in residents of Dong ethnic group in Southeast Guizhou Province

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