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A gold nanoparticles-based lateral flow assay utilizing baculovirus expressed recombinant nucleocapsid and receptor binding domain proteins for serodetection of IgG and IgM against SARS-CoV-2.
Salem, Reda; Elshamy, Amany M; Kamel, Noha; Younes, Soha; Marie, Ohoud M; Waly, Fatma R; El-Kholy, Alaa A; Elmenofy, Wael.
  • Salem R; Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, 12619, Egypt.
  • Elshamy AM; Medical Laboratory Science Department, High Technology Institute of Applied Health Sciences, Badr Academy, Badr City, Cairo, Egypt.
  • Kamel N; Clinical Pathology Department, Faculty of Medicine, Suez Canal University, 4.5 K, Ring Road, Ismailia, 41511, Egypt. noha_mohamed@med.suez.edu.eg.
  • Younes S; Clinical Pathology Department, Faculty of Medicine, Suez Canal University, 4.5 K, Ring Road, Ismailia, 41511, Egypt.
  • Marie OM; Chemistry Department, Faculty of Science, Suez Canal University, Ismailia, 41516, Egypt.
  • Waly FR; Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, 12619, Egypt.
  • El-Kholy AA; Veterinary Sera and Vaccines Research Institute (VSVRI), ARC, Abbassia, P.O. Box # 131, Cairo, 11381, Egypt.
  • Elmenofy W; Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, 12619, Egypt.
Biotechnol Lett ; 44(12): 1507-1517, 2022 Dec.
Article in English | MEDLINE | ID: covidwho-2094663
ABSTRACT
Serological assays for SARS-CoV-2 are being utilized at an exponential rate for surveillance programs. This enterprise was designed to develop and validate a qualitative immunochromatographic test, via the Lateral Flow Assay (LFA), for detection of immunoglobulins M and G (IgM and IgG) against both nucleocapsid (N) and the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2. Both targeted proteins were cloned and expressed in baculovirus expression system utilizing insect cells Sf9. The recombinant RBD and N proteins were purified and conjugated with gold nanoparticles (AuNPs) to set up the coating antigens pad. Both anti-human IgG and IgM were dispensed on nitrocellulose membrane to capture human antibodies in serum samples. A home-made dispensing system was developed to draw identical test and control lines. The validity of the developed LFA was verified by testing serum samples from 103 convalescent COVID-19 patients who were PCR positive for SARS-CoV-2 along with 28 control serum samples. The developed strips showed distinctive bands for IgM and IgG of both proteins (RBD and N) in positive samples. The sensitivity of RBD-based LFA was 70.9% and 39.8% for IgG and IgM, respectively, with a specificity of 100% for both. The N-based LFA exhibited a sensitivity of 73.8% and 35.9% for IgG and IgM, respectively, while its specificity was 75% and 100% for IgG and IgM, respectively. Our developed LFA could afford a tool for surveillance programs in low-resource countries. Moreover, it might be functional for rapid and inexpensive monitoring of the anti-SARS-CoV-2 antibodies in the sera of vaccinated individuals.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Metal Nanoparticles / COVID-19 Type of study: Diagnostic study / Prognostic study / Qualitative research Topics: Vaccines Limits: Humans Language: English Journal: Biotechnol Lett Year: 2022 Document Type: Article Affiliation country: S10529-022-03316-0

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Metal Nanoparticles / COVID-19 Type of study: Diagnostic study / Prognostic study / Qualitative research Topics: Vaccines Limits: Humans Language: English Journal: Biotechnol Lett Year: 2022 Document Type: Article Affiliation country: S10529-022-03316-0