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Evaluation of Three Automated Extraction Systems for the Detection of SARS-CoV-2 from Clinical Respiratory Specimens.
Lim, Ho-Jae; Jung, Hye-Soo; Park, Min-Young; Baek, Young-Hyun; Kannappan, Balaji; Park, Jin-Young; Yang, Jae-Hyun; Seol, Ja-Hwan; Lee, Min-Woo; Jung, Sun-Kyung; Lee, Sun-Hwa; Park, Jung-Eun; Yang, Yong-Jin.
  • Lim HJ; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Jung HS; BK21 FOUR Educational Research Group for Age-Associated Disorder Control Technology, Department of Integrative Biological Sciences, Chosun University, Gwangju 61452, Korea.
  • Park MY; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Baek YH; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Kannappan B; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Park JY; BK21 FOUR Educational Research Group for Age-Associated Disorder Control Technology, Department of Integrative Biological Sciences, Chosun University, Gwangju 61452, Korea.
  • Yang JH; Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA.
  • Seol JH; Paul F. Glenn Center for Biology of Aging Research, Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA.
  • Lee MW; Department of Molecular Medicine, Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA.
  • Jung SK; Department of Integrated Biomedical Science, Soonchunhyang Institute of Medi-Bio Science (SIMS), Soonchunhyang University, Cheonan-si 31151, Korea.
  • Lee SH; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Park JE; Department of Molecular Diagnostics, Seegene Medical Foundation, Seoul 04805, Korea.
  • Yang YJ; BK21 FOUR Educational Research Group for Age-Associated Disorder Control Technology, Department of Integrative Biological Sciences, Chosun University, Gwangju 61452, Korea.
Life (Basel) ; 12(1)2022 Jan 04.
Article in English | MEDLINE | ID: covidwho-1613886
ABSTRACT
Severe acute respiratory syndrome coronavirus (SARS-CoV-2) is highly contagious and causes coronavirus disease 2019 (COVID-19). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is the most accurate and reliable molecular assay to detect active SARS-CoV-2 infection. However, a rapid increase in test subjects has created a global bottleneck in testing capacity. Given that efficient nucleic acid extraction greatly affects reliable and accurate testing results, we compared three extraction platforms MagNA Pure 96 DNA and Viral NA Small Volume kit on MagNA Pure 96 (Roche, Basel, Switzerland), careGENETM Viral/Pathogen HiFi Nucleic Acid Isolation kit (WELLS BIO Inc., Seoul, Korea) on KingFisher Flex (Thermo Fisher Scientific, Rocklin, CA, USA), and SGRespiTM Pure kit (Seegene Inc., Seoul, Korea) on Maelstrom 9600 (Taiwan Advanced Nanotech Inc., Taoyuan, Taiwan). RNA was extracted from 245 residual respiratory specimens from the different types of samples (i.e., NPS, sputum, and saliva) using three different kits. The 95% limits of detection of median tissue culture infectious dose per milliliter (TCID50/mL) for the MagNA Pure 96, KingFisher Flex, and Maelstrom 9600 were 0.37-3.15 × 101, 0.41-3.62 × 101, and 0.33-1.98 × 101, respectively. The KingFisher Flex platform exhibited 99.2% sensitivity and 100% specificity, whereas Maelstrom 9600 exhibited 98.3-100% sensitivity and 100% specificity. Bland-Altman analysis revealed a 95.2% concordance between MagNA Pure 96 and KingFisher Flex and 95.4% concordance between MagNA Pure 96 and Maelstrom 9600, indicating that all three platforms provided statistically reliable results. This suggests that two modifying platforms, KingFisher Flex and Maelstrom 9600, are accurate and scalable extraction platforms for large-scale SARS-CoV-2 clinical detection and could help the management of COVID-19 patients.
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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies / Prognostic study Language: English Year: 2022 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies / Prognostic study Language: English Year: 2022 Document Type: Article