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An encodable multiplex microsphere-phase amplification sensing platform detects SARS-CoV-2 mutations.
Zhong, Zecheng; Wang, Jin; He, Shuizhen; Su, Xiaosong; Huang, Weida; Chen, Mengyuan; Zhuo, Zhihao; Zhu, Xiaomei; Fang, Mujin; Li, Tingdong; Zhang, Shiyin; Ge, Shengxiang; Zhang, Jun; Xia, Ningshao.
  • Zhong Z; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Wang J; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China; School of Life Sciences, Xiamen University, Xiamen, 361102, China.
  • He S; Xiamen Haicang Hospital, Haiyu Road, Xiamen, 361026, China.
  • Su X; Xiang'an Hospital of Xiamen University, Xiang'an East Road, Xiamen, 361102, China.
  • Huang W; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Chen M; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Zhuo Z; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Zhu X; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Fang M; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Li T; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Zhang S; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. Electronic address: zhangshiyin@xmu.edu.cn.
  • Ge S; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. Electronic address: sxge@xmu.edu.cn.
  • Zhang J; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China.
  • Xia N; State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China; School of Life Sciences, Xiamen University, Xiamen, 361102, China; The Researc
Biosens Bioelectron ; 203: 114032, 2022 May 01.
Article in English | MEDLINE | ID: covidwho-1729564
ABSTRACT
SARS-CoV-2 variants of concern (VOCs) contain several single-nucleotide variants (SNVs) at key sites in the receptor-binding region (RBD) that enhance infectivity and transmission, as well as cause immune escape, resulting in an aggravation of the coronavirus disease 2019 (COVID-19) pandemic. Emerging VOCs have sparked the need for a diagnostic method capable of simultaneously monitoring these SNVs. To date, no highly sensitive, efficient clinical tool exists to monitor SNVs simultaneously. Here, an encodable multiplex microsphere-phase amplification (MMPA) sensing platform that combines primer-coded microsphere technology with dual fluorescence decoding strategy to detect SARS-CoV-2 RNA and simultaneously identify 10 key SNVs in the RBD. MMPA limits the amplification refractory mutation system PCR (ARMS-PCR) reaction for specific target sequence to the surface of a microsphere with specific fluorescence coding. This effectively solves the problem of non-specific amplification among primers and probes in multiplex PCR. For signal detection, specific fluorescence codes inside microspheres are used to determine the corresponding relationship between the microspheres and the SNV sites, while the report probes hybridized with PCR products are used to detect the microsphere amplification intensity. The MMPA platform offers a lower SARS-CoV-2 RNA detection limit of 28 copies/reaction, the ability to detect a respiratory pathogen panel without cross-reactivity, and a SNV analysis accuracy level comparable to that of sequencing. Moreover, this super-multiple parallel SNVs detection method enables a timely updating of the panel of detected SNVs that accompanies changing VOCs, and presents a clinical availability that traditional sequencing methods do not.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Topics: Variants Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.bios.2022.114032

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Biosensing Techniques / COVID-19 Type of study: Diagnostic study / Prognostic study / Randomized controlled trials Topics: Variants Limits: Humans Language: English Journal: Biosens Bioelectron Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.bios.2022.114032