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线粒体基因组序列变异作为评估环孢子虫分离株遗传异质性和来源追踪的有用标记。

Mitochondrial genome sequence variation as a useful marker for assessing genetic heterogeneity among Cyclospora cayetanensis isolates and source-tracking.

机构信息

Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, Guangdong, China.

College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450002, Henan, China.

出版信息

Parasit Vectors. 2019 Jan 21;12(1):47. doi: 10.1186/s13071-019-3294-1.

DOI:10.1186/s13071-019-3294-1
PMID:30665345
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6341762/
Abstract

BACKGROUND

Cyclospora cayetanensis is an important enteric pathogen, causing diarrhea and food-borne cyclosporiasis outbreaks. For effective outbreak identification and investigation, it is essential to rapidly assess the genetic heterogeneity of C. cayetanensis specimens from cluster cases and identify the likely occurrence of outbreaks.

METHODS

In this study, we developed a quantitative PCR (qPCR) targeting the polymorphic link region between copies of the mitochondrial genome of C. cayetanensis, and evaluated the genetic heterogeneity among 36 specimens from six countries using melt curve, gel electrophoresis, and sequence analyses of the qPCR products.

RESULTS

All specimens were amplified successfully in the qPCR and produced melt peaks with different Tm values in the melt curve analysis. In gel electrophoresis of the qPCR products, the specimens yielded bands of variable sizes. Nine genotypes were identified by DNA sequencing of the qPCR products. Geographical segregation of genotypes was observed among specimens analyzed, which could be useful in geographical source-tracking.

CONCLUSIONS

The length and nucleotide sequence variations in the mitochondrial genome marker allow rapid assessment of the genetic heterogeneity among C. cayetanensis specimens by melt curve, gel electrophoresis, or DNA sequence analysis of qPCR products. The sequence data generated could be helpful in the initial source-tracking of the pathogen.

摘要

背景

环孢子虫是一种重要的肠道病原体，可引起腹泻和食源性环孢子虫病暴发。为了有效识别和调查暴发，快速评估来自聚集病例的环孢子虫标本的遗传异质性并确定暴发的可能发生至关重要。

方法

本研究中，我们针对环孢子虫线粒体基因组副本之间的多态性连接区开发了一种定量 PCR（qPCR），并使用熔解曲线、凝胶电泳和 qPCR 产物序列分析评估了来自六个国家的 36 个标本的遗传异质性。

结果

所有标本均在 qPCR 中成功扩增，并在熔解曲线分析中产生具有不同 Tm 值的熔解峰。qPCR 产物的凝胶电泳中，标本产生了大小不同的条带。通过 qPCR 产物的 DNA 测序鉴定了 9 种基因型。分析的标本显示基因型存在地理隔离，这有助于进行地理来源追踪。

结论

线粒体基因组标记的长度和核苷酸序列变异允许通过熔解曲线、凝胶电泳或 qPCR 产物的 DNA 序列分析快速评估环孢子虫标本的遗传异质性。生成的序列数据有助于病原体的初步溯源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/7e4cc58309ce/13071_2019_3294_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/4866dc819873/13071_2019_3294_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/fead2a2d05c8/13071_2019_3294_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/7e4cc58309ce/13071_2019_3294_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/4866dc819873/13071_2019_3294_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/fead2a2d05c8/13071_2019_3294_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97d3/6341762/7e4cc58309ce/13071_2019_3294_Fig3_HTML.jpg

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