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PCR-RFLP method to detect zoonotic and host-specific Giardia duodenalis assemblages in dog fecal samples

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Abstract

Giardia duodenalis is a zoonotic protozoan that parasitizes the upper small intestine of human and many mammals including dogs. To develop a restriction fragment length polymorphism (RFLP) method for typing zoonotic (A, B) and host-specific (C, D) assemblages of G. duodenalis from dog, β-giardin gene was amplified with design primer pairs B3 and B4. The PCR products were digested with restriction enzyme Afa I and Msp I; then, PCR-RFLP method was compared with HRM genotyping and sequencing method for G. duodenalis from dog. The results showed that each of assemblages A–D had unique restriction pattern, which was consistent with the predictive results. Among 21 samples tested by PCR-RFLP, 1 human-derived and 8 dog-derived G. duodenalis were identified as assemblage A; 5 dog-derived G. duodenalis as assemblage C; 7 dog-derived G. duodenalis as assemblage D, which were coincided with the HRM genotyping and sequencing results. It is concluded that the PCR-RFLP is quick, easy, and accurate method for the sequence typing of G. duodenalis zoonotic and specific assemblages from dogs.

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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.

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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, Sheng Wu, Auwalu Yusuf Abdullahi, Xinggang Yu, Wei Hu, Meiran Song, Xianli Shi & Guoqing Li

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    1. Liping Tan
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    2. Sheng Wu
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    3. Auwalu Yusuf Abdullahi
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    4. Xinggang Yu
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    8. Guoqing Li
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    Correspondence to Guoqing Li.

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    Liping Tan and Sheng Wu contributed equally to this work.

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    Tan, L., Wu, S., Abdullahi, A.Y. et al. PCR-RFLP method to detect zoonotic and host-specific Giardia duodenalis assemblages in dog fecal samples. Parasitol Res 115, 2045–2050 (2016). https://doi.org/10.1007/s00436-016-4948-y

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    • DOI: https://doi.org/10.1007/s00436-016-4948-y

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