Development of a Novel Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Monkeypox Virus Infections
Chao Yu,
Lulu Zuo,
Jing Miao,
Lingjing Mao,
Benjamin Selekon,
Ella Gonofio,
Emmanuel Nakoune,
Nicolas Berthet,
Gary Wong
Affiliations
Chao Yu
Viral Hemorrhagic Fevers Research Unit, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China
Lulu Zuo
Viral Hemorrhagic Fevers Research Unit, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China
Jing Miao
University of Chinese Academy of Sciences, Beijing 100049, China
Lingjing Mao
University of Chinese Academy of Sciences, Beijing 100049, China
Benjamin Selekon
Laboratory of Arboviruses, Viral Hemorrhagic Fevers, Emerging viruses and Zoonoses, Institut Pasteur of Bangui, Bangui P.O. Box 923, Central African Republic
Ella Gonofio
Laboratory of Arboviruses, Viral Hemorrhagic Fevers, Emerging viruses and Zoonoses, Institut Pasteur of Bangui, Bangui P.O. Box 923, Central African Republic
Emmanuel Nakoune
Laboratory of Arboviruses, Viral Hemorrhagic Fevers, Emerging viruses and Zoonoses, Institut Pasteur of Bangui, Bangui P.O. Box 923, Central African Republic
Nicolas Berthet
Centre for Microbes, Development, and Health, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Unit of Discovery and Molecular Characterization of Pathogens, Shanghai 200031, China
Gary Wong
Viral Hemorrhagic Fevers Research Unit, CAS Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China
A recent outbreak of monkeypox virus (mpox) has prompted researchers to explore diagnostics as a means of impeding transmission and further spread. Rapid, sensitive, and specific methods are crucial for accurately diagnosing mpox infections. Here, we developed a loop-mediated isothermal amplification (LAMP) assay for the specific detection of mpox. The primer sets were designed to target regions in and around the N4R gene, and results showed a detection limit of 2 × 100 DNA copies, which is comparable to the gold-standard qPCR method currently used to detect mpox. Particularly, the assay provides results visible to the naked eye within 30 min. This test specifically detects mpox DNA with no cross-reactivity to related DNA viruses including Varicella Zoster Virus (VZV), Hepatitis B virus (HBV), Vaccinia virus (VACV), Herpes simplex virus-1 (HSV-1), Herpes simplex virus-2 (HSV-2), Human papillomavirus-16 (HPV-16) and Human papillomavirus-18 (HPV-18). Furthermore, the LAMP assay has been evaluated using clinical samples from laboratory-confirmed mpox patients and found to be consistent with the qPCR results. Our results show that this single-tube LAMP method can contribute to diagnosis of suspected mpox infections in the field and clinic, especially in regions with limited laboratory resources.
