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Development of Loop-Mediated Isothermal Amplification (LAMP) for Universal Detection of Enteroviruses

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Abstract

Enteroviruses are found in most environments and cause several diseases in humans. Loop-mediated isothermal amplification (LAMP) was adapted and evaluated for the rapid detection of enteroviruses. Based on the highly conserved 5′ untranslated region (5′-UTR) of the human enteroviruses (HEVs), particularly human enterovirus A (HEV-A) and HEV-B, a set of universal primers was designed. The LAMP amplification was carried out under isothermal conditions at 61 °C, depending on the template concentration results were obtained within 45–90 min. The detection limits were found to be 101 copies of cloned enterovirus 71 fragments, more sensitive than conventional PCR. Nine water samples collected from drinking water sources during three seasons and 19 stool specimens collected from HFMD patients were analyzed. By using the LAMP assay, the majority of samples was tested positive, 9/9 (100 %) and 18/19 (94.7 %), respectively. LAMP is a practical method for the rapid detection of enteroviruses in environmental and clinical samples.

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Acknowledgments

This study was supported by Tianjin Research Program of Application Foundation and Advanced Technology (No.09JCYBJC08400 and No.10JCZDJC24600).

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Authors and Affiliations

  1. Tianjin Key Laboratory for Biomarkers of Occupational and Environmental Hazard, Logistics University of Chinese People’s Armed Police Forces, Tianjin, 300162, China

    Hua-Bing Zhao, Guang-Ya Yin, Guo-Ping Zhao, Ai-Hua Huang, Jun-Hong Wang, Shuang-Feng Yang & Hong-Sheng Gao

  2. School of Public Health, Hebei Medical University, Shijiazhuang, 050031, China

    Wei-Jun Kang

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  1. Hua-Bing Zhao
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  2. Guang-Ya Yin
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  3. Guo-Ping Zhao
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Correspondence to Hua-Bing Zhao.

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Zhao, HB., Yin, GY., Zhao, GP. et al. Development of Loop-Mediated Isothermal Amplification (LAMP) for Universal Detection of Enteroviruses. Indian J Microbiol 54, 80–86 (2014). https://doi.org/10.1007/s12088-013-0399-7

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  • DOI: https://doi.org/10.1007/s12088-013-0399-7

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