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A simple and efficient method for potential point-of-care diagnosis of human papillomavirus genotypes: combination of isothermal recombinase polymerase amplification with lateral flow dipstick and reverse dot blot

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Abstract

Cervical cancer is the second most common cancer in the world’s woman population with a high incidence in developing countries where diagnostic conditions for the cancer are poor. The main culprit causing the cancer is the human papillomavirus (HPV). HPV is divided into three major groups, i.e., high-risk (HR) group, probable high-risk (pHR) group, and low-risk (LR) group according to their potential of causing cervical cancer. Therefore, developing a sensitive, reliable, and cost-effective point-of-care diagnostic method for the virus genotypes in developing countries even worldwide is of high importance for the cancer prevention and control strategies. Here we present a combined method of isothermal recombinase polymerase amplification (RPA), lateral flow dipstick (LFD), and reverse dot blot (RDB), in quick point-of-care identification of HPV genotypes. The combined method is highly specific to HPV when the conserved L1 genes are used as targeted genes for amplification. The method can be used in identification of HPV genotypes at point-of-care within 1 h with a sensitivity of low to 100 fg of the virus genomic DNA. We have demonstrated that it is an excellent diagnostic point-of-care assay in monitoring the disease without time-consuming and expensive procedures and devices.

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Acknowledgments

We thank Wenzhou People’s Hospital and Mei Zhong Medical Laboratory for providing the cervical scrape samples.

Funding

The work was supported by the National Key Research and Development Program of China (2017YFF0210200, 2018YFF0215205) and the Public Projects of Zhejiang Province (LGC19C200006).

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Authors and Affiliations

  1. Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou, 310018, Zhejiang, China

    Biao Ma, Jiehong Fang, Wei Lin, Xiaoping Yu & Mingzhou Zhang

  2. Department of Plant Biology, Uppsala BioCenter, Linnean Centre for Plant Biology, Swedish University of Agricultural Science (SLU), P.O. Box 7080, SE-75007, Uppsala, Sweden

    Chuanxin Sun

Authors
  1. Biao Ma
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  2. Jiehong Fang
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  3. Wei Lin
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  4. Xiaoping Yu
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  5. Chuanxin Sun
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  6. Mingzhou Zhang
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Correspondence to Chuanxin Sun or Mingzhou Zhang.

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The ethical committee of Wenzhou People’s Hospital approved the study.

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Ma, B., Fang, J., Lin, W. et al. A simple and efficient method for potential point-of-care diagnosis of human papillomavirus genotypes: combination of isothermal recombinase polymerase amplification with lateral flow dipstick and reverse dot blot. Anal Bioanal Chem 411, 7451–7460 (2019). https://doi.org/10.1007/s00216-019-02113-5

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