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在中国,使用环介导等温扩增(LAMP)和聚合酶链式反应(PCR)技术从天然水库、污水和制药厂污水中检测棘阿米巴属。

Detection of Acanthamoeba spp. in water samples collected from natural water reservoirs, sewages, and pharmaceutical factory drains using LAMP and PCR in China.

机构信息

State Key Laboratory of Plateau Ecology and Agriculture, Center for Biomedicine and Infectious Disease, Qinghai Academy of Animal Sciences and Veterinary Medicine, Medical School of Qinghai University Xining, 1#Wei'er Road, Qinghai, Biological Scientific Estate Garden, Xining 810016, PR China; Department of Tropical Parasitology, Institute of Maritime and Tropical Medicine in Gdynia, Medical University of Gdansk, 9b Powstania Styczniowego Str, 81-519 Gdynia, Poland.

State Key Laboratory of Plateau Ecology and Agriculture, Center for Biomedicine and Infectious Disease, Qinghai Academy of Animal Sciences and Veterinary Medicine, Medical School of Qinghai University Xining, 1#Wei'er Road, Qinghai, Biological Scientific Estate Garden, Xining 810016, PR China; Research Group of Functional Materials and Catalysis, GIMFC, Universidad de Nariño, Calle 18 Cra. 50, Campus Torobajo, Pasto-Colombia. Doctorate in Biotechnology, Universidad Nacional de Colombia, Bogotá D.C., Colombia.

出版信息

Sci Total Environ. 2017 Apr 15;584-585:489-494. doi: 10.1016/j.scitotenv.2017.01.046. Epub 2017 Jan 26.

Abstract

Various species of amoebas belonging to the genus Acanthamoeba are widely distributed in many parts of the world. Some strains of these protozoans may exist as parasites and pose risks to human health as causative agents of serious human diseases. Currently in China there is a lack of information about the distribution of Acanthamoeba strains in the environment. Accordingly, 261 environmental water samples taken from rivers, sewage, and pharmaceutical factory drains were collected in Qinghai Province, China. The material was filtered and then analysed with both LAMP and PCR assays. Of the samples examined, Acanthamoeba DNA was found in 32 (14.68%) samples with the use of LAMP; in 13 of these samples, DNA from this amoeba was also detected using PCR. Sequencing of selected positive samples confirmed that the PCR products were fragments of the Acanthamoeba 18S rRNA gene and that isolates represent the T4 genotype, known as the most common strain related to AK cases. The results indicate that surface water, as well as water taken from sewage and pharmaceutical drains, may be a source of acanthamoebic strains potentially pathogenic for humans in China. It has been also demonstrated that LAMP assays is more sensitive than PCR and can be regarded as useful tool for screening the environment for Acanthamoeba spp.

摘要

属于棘阿米巴属的各种变形虫广泛分布于世界许多地区。这些原生动物的一些菌株可能以寄生虫的形式存在,并作为严重人类疾病的病原体对人类健康构成威胁。目前,中国对环境中棘阿米巴菌株的分布情况知之甚少。因此,从中国青海省的河流、污水和制药厂排水中采集了 261 个环境水样。对采集的水样进行过滤,然后用 LAMP 和 PCR 检测。使用 LAMP 检测到 32 个(14.68%)样本中存在棘阿米巴 DNA;在其中 13 个样本中,也使用 PCR 检测到了这种变形虫的 DNA。对选定的阳性样本进行测序证实,PCR 产物是棘阿米巴 18S rRNA 基因的片段,分离株代表 T4 基因型,被认为是与 AK 病例相关的最常见菌株。结果表明,地表水以及污水和制药厂排水可能是中国潜在致病性棘阿米巴菌株的来源。研究还表明,LAMP 检测比 PCR 更敏感,可作为筛选环境中棘阿米巴属的有用工具。

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