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Establishment and preliminary application of nanoparticle-assisted PCR assay for detection of Cryptosporidium spp.

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Abstract

Cryptosporidium is an important intestinal protozoan parasite that causes diarrhoea in humans and animals. To rapidly and specifically detect Cryptosporidium spp., we designed a pair of primers based on the small subunit ribosomal RNA (SSU rRNA) gene of Cryptosporidium spp. to be used in a new nanoparticle-assisted PCR (nano-PCR) assay. The minimum detectable concentration (1.02 pg) of this nano-PCR was 10 times more sensitive than conventional PCR using the same primer pair. The DNA samples of C. parvum, C. baileyi, C. xiaoi, C. ryanae, and C. andersoni were successfully detected by the nano-PCR. No amplifications were evident with DNA samples of some common intestinal pathogens, including Eimeria tenella, Blastocystis sp., Giardia lamblia, Enterocytozoon bieneusi, and Balantidium coli. To validate the clinical usefulness of the novel nano-PCR, a total of 40 faecal samples from goats, camels, calves, and chickens were examined. The positive rate of Cryptosporidium spp. was 27.5% (11/40), which was consistent with that of an established nested PCR. These results indicate that the novel nano-PCR assay enables the rapid, specific, and accurate detection of Cryptosporidium infection in animals. The findings provide a technical basis for the clinical diagnosis, prevention, and control of cryptosporidiosis.

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Funding

This study was funded by the grants from the National Key Research and Development Program of China (Grant No. 2017YFD0501305), National Natural Science Foundation of China (32072890), and the Open Funds of the State Key Laboratory of Veterinary Aetiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences (SKLVEB2020KFKT015).

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Author notes

  1. Yan-Ling Yin and Yi Wang contributed equally to this work.

Authors and Affiliations

  1. College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi Province, People’s Republic of China

    Yan-Ling Yin, Yi Wang, Peng Lai, Qian Yao, Yuan Li, Xin Yang, Jun-Ke Song & Guang-Hui Zhao

  2. College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, 450046, Henan Province, People’s Republic of China

    Long-Xian Zhang

Authors
  1. Yan-Ling Yin
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  2. Yi Wang
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  5. Yuan Li
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  9. Guang-Hui Zhao
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Correspondence to Jun-Ke Song or Guang-Hui Zhao.

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Ethics approval

Our research was conducted strictly in accordance with the Guide for the Care and Use of Laboratory Animals of the Ministry of Health, China, and the protocol was reviewed and approved by the Research Ethics Committee of Northwest A&F University, Yangling, China. Permission was obtained from animal owners prior to faecal collection.

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The authors declare no competing interests.

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Section Editor: Lihua Xiao

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Yin, YL., Wang, Y., Lai, P. et al. Establishment and preliminary application of nanoparticle-assisted PCR assay for detection of Cryptosporidium spp.. Parasitol Res 120, 1837–1844 (2021). https://doi.org/10.1007/s00436-021-07101-2

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