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Sensitive and rapid detection of Paragonimus westermani infection in humans and animals by loop-mediated isothermal amplification (LAMP)

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Abstract

In the present study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the detection of Paragonimus westermani adults, metacercariae, and eggs in human and animal samples. The LAMP amplification can be finished in 45 min under isothermal condition at 60°C by employing a set of four species-specific primer mixtures and the results can be checked by naked-eye visualization. No amplification products were detected with deoxyribunucleic acid (DNA) of related trematode species including Fasciola hepatica, Fasciola gigantica, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma mansoni, and Schistosoma japonicum. The method was further validated by examining P. westermani DNA in intermediate hosts including freshwater crabs and crayfish, as well as in sputum and pleural fluid samples from patients of paragonimiasis. These results indicated that the LAMP assay was highly specific, sensitive, and rapid, and it was approximately 100 times more sensitive than conventional specific PCR. The LAMP assay established in this study provides a rapid and sensitive tool for the detection of P. westermani DNA in freshwater crabs, crayfish, sputum, and pleural fluid samples, which has important implications for effective control of human paragonimiasis.

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Acknowledgements

This work is supported in part by the Program for National S&T Major Program (grant no. 2008ZX10004-011), National Key Technology R&D Program (grant no. 2008BAI56B03) and the State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The experiments comply with the current laws of the country in which the experiments were performed.

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Authors and Affiliations

  1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, 200025, People’s Republic of China

    M. X. Chen, L. Ai, S. H. Chen, Y. N. Zhang & J. X. Chen

  2. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People’s Republic of China

    M. J. Xu & X. Q. Zhu

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

    L. Ai & X. Li

  4. Shenzhen Center for Disease Control and Prevention, Shenzhen, Guangdong Province, 518020, People’s Republic of China

    R. L. Zhang

  5. Shenzhen Medical Continuing Education Center, Shenzhen, Guangdong Province, 518020, People’s Republic of China

    J. J. Xia & K. Wang

Authors
  1. M. X. Chen
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  2. L. Ai
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  3. R. L. Zhang
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  4. J. J. Xia
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  5. K. Wang
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  6. S. H. Chen
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  7. Y. N. Zhang
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  8. M. J. Xu
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  9. X. Li
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  10. X. Q. Zhu
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  11. J. X. Chen
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Corresponding authors

Correspondence to X. Q. Zhu or J. X. Chen.

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M. X. Chen and L. Ai contributed equally to this work.

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Chen, M.X., Ai, L., Zhang, R.L. et al. Sensitive and rapid detection of Paragonimus westermani infection in humans and animals by loop-mediated isothermal amplification (LAMP). Parasitol Res 108, 1193–1198 (2011). https://doi.org/10.1007/s00436-010-2162-x

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  • DOI: https://doi.org/10.1007/s00436-010-2162-x

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