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Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia

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Abstract

Giardia lamblia is a zoonotic flagellate protozoan in the intestine of human and many mammals including dogs. To assess a threat of dog-derived G. lamblia to humans, the common dog-derived G. lamblia assemblages A, C, and D were genotyped by high-resolution melting (HRM) technology. According to β-giardin gene sequence, the qPCR-HRM primers BG5 and BG7 were designed. A series of experiments on the stability, sensitivity, and accuracy of the HRM method were also tested. Results showed that the primers BG5 and BG7 could distinguish among three assemblages A, C, and D, which Tm value differences were about 1 °C to each other. The melting curves of intra-assay reproducibility were almost coincided, and those of inter-assay reproducibility were much the same shape. The lowest detection concentration was about 5 × 10−6-ng/μL sample. The genotyping results from 21 G. lamblia samples by the HRM method were in complete accordance with sequencing results. It is concluded that the HRM genotyping method is rapid, stable, specific, highly sensitive, and suitable for clinical detection and molecular epidemiological survey of dog-derived G. lamblia.

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Acknowledgments

This work was funded by the National Natural Science Foundation of China (Grant no. 31272551). The authors would like to thank the humane shelter’s personnel for helping to collect clinical samples and Miss Ping Kang for helping to draft the manuscript.

Conflict of interests

The authors declare that there is no conflict of interests regarding the publication of this paper.

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Authors and Affiliations

  1. College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People’s Republic of China

    Liping Tan, Xingang Yu, Auwalu Yusuf Abdullahi, Sheng Wu, Guochao Zheng, Wei Hu, Meiran Song, Zhen Wang, Biao Jiang & Guoqing Li

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  1. Liping Tan
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  2. Xingang Yu
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  3. Auwalu Yusuf Abdullahi
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  4. Sheng Wu
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  5. Guochao Zheng
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  6. Wei Hu
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  7. Meiran Song
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  8. Zhen Wang
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  9. Biao Jiang
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  10. Guoqing Li
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Correspondence to Guoqing Li.

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Tan, L., Yu, X., Abdullahi, A.Y. et al. Development of a rapid HRM genotyping method for detection of dog-derived Giardia lamblia . Parasitol Res 114, 4081–4086 (2015). https://doi.org/10.1007/s00436-015-4636-3

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  • DOI: https://doi.org/10.1007/s00436-015-4636-3

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