慢性腹泻患者艰难梭菌、人芽囊原虫和隐孢子虫感染的研究

doi:10.12140/j.issn.1000-7423.2019.06.020

中国寄生虫学与寄生虫病杂志 ›› 2019, Vol. 37 ›› Issue (6): 723-726.doi: 10.12140/j.issn.1000-7423.2019.06.020

慢性腹泻患者艰难梭菌、人芽囊原虫和隐孢子虫感染的研究

周亚玲¹(), 吴亮², 阴晴^1,^*(), 吴瑶¹, 何蕾¹, 邹治情², 戴晓玥², 夏雯²

1 江苏大学附属医院检验科,镇江 212001
2 江苏大学医学院,镇江 212013

收稿日期:2019-06-06 出版日期:2019-12-30 发布日期:2019-12-31
通讯作者: 阴晴
作者简介:
作者简介：周亚玲（1993-）,女,硕士,从事微生物方向研究。E-mail: 2691891576@qq.com
基金资助:
镇江市2017年度科技创新资金（重点研发计划-社会发展项目）（No. SH2017024）;江苏省2018年度预防医学科研课题（No. Y2018108）;中国科技部基础平台项目（No. 平台-TDRC-22）

Infections of Clostridium difficile, Blastocystis hominis and Cryptosporidium spp. in patients with chronic diarrhea

Ya-ling ZHOU¹(), Liang WU^2,^*, Qing YIN^1,^*(), Yao WU¹, Lei HE¹, Zhi-qing ZOU², Xiao-yue DAI², Wen XIA²

1 The Department of Clinical Laboratory, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China
2 School of Medicine, Jiangsu University, Zhenjiang 212013, China

Received:2019-06-06 Online:2019-12-30 Published:2019-12-31
Contact: Liang WU,Qing YIN
Supported by:
Supported by 2017 Science and Technology Innovation Fund of Zhenjiang City(No. SH2017024), 2018 Preventive Medicine Research Project of Jiangsu Province(No. Y2018108), Basic Platform Project of the Ministry of Science and Technology of China(No. platform-TDRC-22)

摘要/Abstract

摘要：

收集江苏大学附属医院2018年收治的慢性腹泻患者粪样,涂片镜检并提取粪样DNA,采用多重PCR技术检测粪样中艰难梭菌16S rDNA基因、毒素A（tcdA）基因、毒素B（tcdB）基因和二元毒素（cdtA和cdtB）基因;常规PCR检测人芽囊原虫SSU rRNA基因;巢式PCR检测隐孢子虫SSU rRNA基因。96份慢性腹泻患者粪样中,检出艰难梭菌16S rDNA基因64份,阳性率为66.7%;艰难梭菌阳性粪样中,tcdA基因24例,tcdB基因18例,未检出单独携带tcdB基因粪样,产毒株携带率为37.5%,所有粪样中均未检出cdtA基因和cdtB基因;镜检结果显示,人芽囊原虫粪样16份,检出率为16.7%;PCR结果显示,21份粪样扩增出人芽囊原虫SSU rRNA基因,检出率为21.9%,镜检阳性粪样PCR检测也为阳性;未检测到艰难梭菌和人芽囊原虫混合感染;未检出隐孢子虫SSU rRNA基因。江苏大学附属医院慢性腹泻患者中主要检出的病原体为艰难梭菌和人芽囊原虫。

关键词: 慢性腹泻, 艰难梭菌, 人芽囊原虫, 隐孢子虫

Abstract:

Ninety-six stool samples were collected from patients with chronic diarrhea at affiliated hospital of Jiangsu University (inpatients and outpatients) in 2018. Each stool sample was smeared for microscopic examination. At the same time fecal DNA was extracted. The genes for 16S rDNA, toxin A (tcdA), toxin B (tcdB), binary toxin (cdtA and cdtB) of Clostridium difficile were detected by multiplex PCR. The SSU rRNA gene of Blastocystis hominis was detected by conventional PCR and the SSU rRNA gene of Cryptosporidium spp. was detected by nest-PCR. C. difficile 16S rDNA was detected in 64 out of 96 patients with chronic diarrhea with a positive rate of 66.7%. C. difficile TcdA gene was detected in 24 cases and tcdB gene in 18 cases of all C. difficile positive samples. There was no sample with only tcdB. No cdtA and cdtB genes were detected in any C. difficile positive sample. The rate of toxigenic strain was 37.5%. Microscopic examination detected 16 B. hominis infected samples with detection rate of 16.7%. PCR results showed that 21 samples B. hominis SSU rRNA gene was amplified in 96 stool samples with a detection rate of 21.9%. All microscopic positive samples were positive in PCR detection. No mixed infection of C. difficile and B. hominis was detected. The Cryptosporidium spp. SSU rRNA gene was not detected in all stool samples. The results showed that C. difficile and B. hominis were the main organisms detected in patients with chronic diarrhea in Nanjing area.

Key words: Chronic diarrhea, Clostridium difficile, Blastocystis hominis, Cryptosporidium spp.

中图分类号:

R532.23

周亚玲, 吴亮, 阴晴, 吴瑶, 何蕾, 邹治情, 戴晓玥, 夏雯. 慢性腹泻患者艰难梭菌、人芽囊原虫和隐孢子虫感染的研究[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(6): 723-726.

Ya-ling ZHOU, Liang WU, Qing YIN, Yao WU, Lei HE, Zhi-qing ZOU, Xiao-yue DAI, Wen XIA. Infections of Clostridium difficile, Blastocystis hominis and Cryptosporidium spp. in patients with chronic diarrhea[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2019, 37(6): 723-726.

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基因	引物序列（5′→3′）	预计产物大小/bp
艰难梭菌 tcdA	GCATGATAAGGCAACTTCAGTGGTA	629
	AGTTCCTCCTGCTCCATCAAATG
tcdB	CCAAARTGGAGTGTTACAAACAGGTG
	GCATTTCTCCATTCTCAGCAAAGTA	410
	GCATTTCTCCGTTTTCAGCAAAGTA
cdtA	GGGAAGCACTATATTAAAGCAGAAGC
	GGGAAACATTATATTAAAGCAGAAGC	221
	CTGGGTTAGGATTATTTACTGGACCA
cdtB	TTGACCCAAAGTTGATGTCTGATTG	262
	CGGATCTCTTGCTTCAGTCTTTATAG
16S rDNA	GGAGGCAGCAGTGGGGAATA	1 062
	TGACGGGCGGTGTGTACAAG
人芽囊原虫 SSU rRNA	GAGCTTTTTAAGTCCAACAACG	600
	ATCTGGTTCATGGTTGCGAGT
隐孢子虫 SSUrRNA 第1轮	TTCTAGAGCTAATACATGCG	-
	CCCATTTCCTTCGAAACAGGA
第2轮	GGAAGGGTTGTATTTATTAGATAAAG	850
	AAGGAGTAAGGAACAACCTCCA

慢性腹泻患者艰难梭菌、人芽囊原虫和隐孢子虫感染的研究

Infections of Clostridium difficile, Blastocystis hominis and Cryptosporidium spp. in patients with chronic diarrhea

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