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Matrix-Assisted Laser Desorption and Ionization Time-of-Flight Mass Spectrometry Analysis for the Direct Detection of SARS-CoV-2 in Nasopharyngeal Swabs.
Yoshinari, Tomoya; Hayashi, Katsuhiko; Hirose, Shouhei; Ohya, Kenji; Ohnishi, Takahiro; Watanabe, Maiko; Taharaguchi, Satoshi; Mekata, Hirohisa; Taniguchi, Takahide; Maeda, Takuya; Orihara, Yuta; Kawamura, Rieko; Arai, Sakura; Saito, Yoshiro; Goda, Yukihiro; Hara-Kudo, Yukiko.
  • Yoshinari T; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Hayashi K; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Hirose S; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Ohya K; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Ohnishi T; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Watanabe M; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Taharaguchi S; Laboratory of Microbiology, Department of Veterinary Medicine, Azabu University, 1-17-71 Fucihnobe, Chuo-ku, Sagamihara, Kanagawa 252-5201, Japan.
  • Mekata H; Center for Animal Disease Control, University of Miyazaki, 1-1 Gakuen Kibanadai-nishi, Miyazaki 889-2192, Japan.
  • Taniguchi T; Division of Animal Life Science, Institute of Agriculture, Tokyo University of Agriculture and Technology, 3-5-8 Saiwai-cho, Fuchu-shi, Tokyo 183-8509, Japan.
  • Maeda T; Department of Clinical Laboratory, Saitama Medical University Hospital, 38 Morohongo Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan.
  • Orihara Y; Department of Clinical Laboratory, Saitama Medical University Hospital, 38 Morohongo Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan.
  • Kawamura R; Department of Clinical Laboratory, Saitama Medical University Hospital, 38 Morohongo Moroyama-machi, Iruma-gun, Saitama 350-0495, Japan.
  • Arai S; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Saito Y; Division of Medicinal Safety Science, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Goda Y; Director General, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
  • Hara-Kudo Y; Division of Microbiology, National Institute of Health Sciences, 3-25-26 Tonomachi, Kawasaki-ku, Kawasaki City, Kanagawa 210-9501, Japan.
Anal Chem ; 94(10): 4218-4226, 2022 03 15.
Article in English | MEDLINE | ID: covidwho-1721377
ABSTRACT
The most common diagnostic method used for coronavirus disease-2019 (COVID-19) is real-time reverse transcription polymerase chain reaction (PCR). However, it requires complex and labor-intensive procedures and involves excessive positive results derived from viral debris. We developed a method for the direct detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs, which uses matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-ToF MS) to identify specific peptides from the SARS-CoV-2 nucleocapsid phosphoprotein (NP). SARS-CoV-2 viral particles were separated from biological molecules in nasopharyngeal swabs by an ultrafiltration cartridge. Further purification was performed by an anion exchange resin, and purified NP was digested into peptides using trypsin. The peptides from SARS-CoV-2 that were inoculated into nasopharyngeal swabs were detected by MALDI-ToF MS, and the limit of detection was 106.7 viral copies. This value equates to 107.9 viral copies per swab and is approximately equivalent to the viral load of contagious patients. Seven NP-derived peptides were selected as the target molecules for the detection of SARS-CoV-2 in clinical specimens. The method detected between two and seven NP-derived peptides in 19 nasopharyngeal swab specimens from contagious COVID-19 patients. These peptides were not detected in four specimens in which SARS-CoV-2 RNA was not detected by PCR. Mutated NP-derived peptides were found in some specimens, and their patterns of amino acid replacement were estimated by accurate mass. Our results provide evidence that the developed MALDI-ToF MS-based method in a combination of straightforward purification steps and a rapid detection step directly detect SARS-CoV-2-specific peptides in nasopharyngeal swabs and can be a reliable high-throughput diagnostic method for COVID-19.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Anal Chem Year: 2022 Document Type: Article Affiliation country: Acs.analchem.1c04328

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Anal Chem Year: 2022 Document Type: Article Affiliation country: Acs.analchem.1c04328