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Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces

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Abstract

Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.

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Acknowledgments

Project support was provided by the National Key Project of Scientific and Technical Supporting Program (Grant No. 2012BAD12B07), the Special Fund for Agro-scientific Research in the Public Interest (Grant No. 201303037), and the Science and Technology Support Program of Gansu Province (Grant No. 1504NKCA054-6).

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Authors and Affiliations

  1. 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

    Yao-Dong Wu, Dong-Hui Zhou, Wen-Bin Zheng, Jian-Gang Ma, Meng Wang, Xing-Quan Zhu & Min-Jun Xu

  2. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province, 450002, People’s Republic of China

    Long-Xian Zhang

  3. College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province, 130118, People’s Republic of China

    Wen-Bin Zheng & Jian-Gang Ma

  4. College of Animal Science, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People’s Republic of China

    Min-Jun Xu

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  1. Yao-Dong Wu
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  2. Dong-Hui Zhou
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  3. Long-Xian Zhang
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  7. Xing-Quan Zhu
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Correspondence to Min-Jun Xu.

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Wu, YD., Zhou, DH., Zhang, LX. et al. Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces. Parasitol Res 115, 3551–3555 (2016). https://doi.org/10.1007/s00436-016-5120-4

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