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Aptamer/antibody sandwich method for digital detection of SARS-CoV2 nucleocapsid protein.
Ge, Chenchen; Feng, Juan; Zhang, Jiaming; Hu, Kai; Wang, Dou; Zha, Ling; Hu, Xuejuan; Li, Rongsong.
  • Ge C; College of Health Science and Environmental Engineering, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China.
  • Feng J; College of Health Science and Environmental Engineering, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China.
  • Zhang J; Sino-German College of Intelligent Manufacturing, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China.
  • Hu K; Sino-German College of Intelligent Manufacturing, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China.
  • Wang D; Shenzhen Key Laboratory of Smart Healthcare Engineering, Department of Biomedical Engineering, Southern University of Science and Technology, No. 1088, Xueyuan Rd., Xili, Nanshan District, Shenzhen, Guangdong, 518055, PR China. Electronic address: wangd9@sustech.edu.cn.
  • Zha L; College of Health Science and Environmental Engineering, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China.
  • Hu X; Sino-German College of Intelligent Manufacturing, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China. Electronic address: huxuejuan@sztu.edu.cn.
  • Li R; College of Health Science and Environmental Engineering, Shenzhen Technology University, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong, 518118, PR China. Electronic address: lirongsong@sztu.edu.cn.
Talanta ; 236: 122847, 2022 Jan 01.
Article in English | MEDLINE | ID: covidwho-1401881
ABSTRACT
Nucleocapsid protein (N protein) is the most abundant protein in SARS-CoV2 and is highly conserved, and there are no homologous proteins in the human body, making it an ideal biomarker for the early diagnosis of SARS-CoV2. However, early detection of clinical specimens for SARS-CoV2 remains a challenge due to false-negative results with viral RNA and host antibodies based testing. In this manuscript, a microfluidic chip with femtoliter-sized wells was fabricated for the sensitive digital detection of N protein. Briefly, ß-galactosidase (ß-Gal)-linked antibody/N protein/aptamer immunocomplexes were formed on magnetic beads (MBs). Afterwards, the MBs and ß-Gal substrate fluorescein-di-ß-d-galactopyranoside (FDG) were injected into the chip together. Each well of the chip would only hold one MB as confined by the diameter of the wells. The MBs in the wells were sealed by fluorocarbon oil, which confines the fluorescent (FL) product generated from the reaction between ß-Gal and FDG in the individual femtoliter-sized well and creates a locally high concentration of the FL product. The FL images of the wells were acquired using a conventional inverted FL microscope. The number of FL wells with MBs (FL wells number) and the number of wells with MBs (MBs wells number) were counted, respectively. The percentage of FL wells was calculated by dividing (FL wells number) by (MBs wells number). The higher the percentage of FL wells, the higher the N protein concentration. The detection limit of this digital method for N protein was 33.28 pg/mL, which was 300 times lower than traditional double-antibody sandwich based enzyme-linked immunosorbent assay (ELISA).
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoassay / Nucleocapsid Proteins / SARS-CoV-2 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Talanta Year: 2022 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoassay / Nucleocapsid Proteins / SARS-CoV-2 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Talanta Year: 2022 Document Type: Article