title: Easy-to-use rapid gene amplification method for direct detection of RNA and DNA viruses in sera and feces from various animals.
abstract: The development of rapid and simple gene amplification tests is required for detection of pathogens to prevent transmission of infectious diseases between animals or from animals to humans. An easy-to-use rapid gene amplification method that can directly detect RNA and DNA viruses in clinical samples was developed. This method is based on combining loop-mediated isothermal amplification (LAMP) or reverse transcription-LAMP (RT-LAMP) and RNA GEM Tissue, a thermophilic enzyme that extracts nucleic acid by quickly digesting proteins and ribonucleases. The authors named these methods GEM LAMP and GEM RT-LAMP. These methods were able to detect viral DNA and RNA within 70 min in a single tube using only a water bath. The detection capacities were 10-100-fold more sensitive than those of previously established LAMP and RT-LAMP methods. The GEM LAMP and GEM RT-LAMP methods were used to detect macroscopically the presence of DNA and RNA viruses in sera or fecal samples from cattle, pigs, horses, dolphins, penguins, and sea lions using SYBR green I. The GEM LAMP and GEM RT-LAMP methods thus have considerable versatility as tools for detecting pathogens and are applicable to basic human and veterinary medicine, environmental hygiene, and point-of-care-testing.
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Segawa
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kobayashi
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Yukina
authors.initials: Y
authors.lastname: Sase
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan. Electronic address: itou.takuya@nihon-u.ac.jp.
authors.forename: Takuya
authors.initials: T
authors.lastname: Itou
authors.affiliation: Department of Marine Science and Resources, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Miwa
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Shinagawa Aquarium, 3-2-1 Katsushima, Shinagawa-ku, Tokyo 140-0012, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Endoh
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nakanishi
authors.affiliation: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Sakai
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D003959:Diamines; D005243:Feces / Q000821:virology*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D044967:Serum / Q000821:virology*; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors; D014777:Virus Diseases / Q000175:diagnosis* / Q000662:veterinary* / Q000821:virology
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
keywords: Direct detection; LAMP; RNA and DNA viruses; easy-to-use