tetano
Editor, Senior Moderator
Sci Rep
. 2024 Sep 17;14(1):21655.
doi: 10.1038/s41598-024-72453-5. Monitoring SARS-CoV-2 IgA, IgM and IgG antibodies in dried blood and saliva samples using antibody proximity extension assays (AbPEA)
Mengqi Wang[SUP] 1 [/SUP], Masood Kamali-Moghaddam[SUP] 1 2 [/SUP], Liza Löf[SUP] 1 2 [/SUP], Matilde Cortabarría Fernandez[SUP] 1 3 [/SUP], Roger Díaz Codina[SUP] 1 [/SUP], Fredrik H Sterky[SUP] 4 5 6 [/SUP], Mikael Åberg[SUP] 2 7 [/SUP], Ulf Landegren[SUP] 8 [/SUP], Hongxing Zhao[SUP] 9 10 [/SUP]
Affiliations
Using a modified proximity extension assay, total and immunoglobulin (Ig) class-specific anti-SARS-CoV-2 antibodies were sensitively and conveniently detected directly from ø1.2 mm discs cut from dried blood and saliva spots (DBS and DSS) without the need for elution. For total Ig detection, antigen probes were prepared by conjugating recombinant spike protein subunit 1 (S1-RBD) to a pair of oligonucleotides. To detect isotype-specific antibody reactivity, one antigen probe was replaced with oligonucleotide-conjugated antibodies specific for antibody isotypes. Binding of pairs of oligonucleotide-conjugated probes to antibodies in patient samples brings oligonucleotides in proximity. An added DNA polymerase uses a transient hybridization between the oligonucleotides to prime synthesis of a DNA strand, which serves as a DNA amplicon that is quantified by real-time PCR. The S1-RBD-specific IgG, IgM, and IgA antibodies in DBS samples collected over the course of a first and second vaccination exhibited kinetics consistent with previous reports. Both DBS and DSS collected from 42 individuals in the autumn of 2023 showed significant level of total S1-RBD antibodies with a correlation of R = 0.70. However, levels in DSS were generally 10 to 100-fold lower than in DBS. Anti-S1-RBD IgG and IgA in DSS demonstrated a correlation of R = 0.6.
Keywords: Antibody isotypes; Antibody proximity extension assay; Dried blood spot (DBS); Dried saliva spot (DSS); IgA; IgG; IgM; Immunoassays; Real-time PCR; SARS-CoV-2 antibody; Vaccination.
. 2024 Sep 17;14(1):21655.
doi: 10.1038/s41598-024-72453-5. Monitoring SARS-CoV-2 IgA, IgM and IgG antibodies in dried blood and saliva samples using antibody proximity extension assays (AbPEA)
Mengqi Wang[SUP] 1 [/SUP], Masood Kamali-Moghaddam[SUP] 1 2 [/SUP], Liza Löf[SUP] 1 2 [/SUP], Matilde Cortabarría Fernandez[SUP] 1 3 [/SUP], Roger Díaz Codina[SUP] 1 [/SUP], Fredrik H Sterky[SUP] 4 5 6 [/SUP], Mikael Åberg[SUP] 2 7 [/SUP], Ulf Landegren[SUP] 8 [/SUP], Hongxing Zhao[SUP] 9 10 [/SUP]
Affiliations
- PMID: 39289450
- DOI: 10.1038/s41598-024-72453-5
Using a modified proximity extension assay, total and immunoglobulin (Ig) class-specific anti-SARS-CoV-2 antibodies were sensitively and conveniently detected directly from ø1.2 mm discs cut from dried blood and saliva spots (DBS and DSS) without the need for elution. For total Ig detection, antigen probes were prepared by conjugating recombinant spike protein subunit 1 (S1-RBD) to a pair of oligonucleotides. To detect isotype-specific antibody reactivity, one antigen probe was replaced with oligonucleotide-conjugated antibodies specific for antibody isotypes. Binding of pairs of oligonucleotide-conjugated probes to antibodies in patient samples brings oligonucleotides in proximity. An added DNA polymerase uses a transient hybridization between the oligonucleotides to prime synthesis of a DNA strand, which serves as a DNA amplicon that is quantified by real-time PCR. The S1-RBD-specific IgG, IgM, and IgA antibodies in DBS samples collected over the course of a first and second vaccination exhibited kinetics consistent with previous reports. Both DBS and DSS collected from 42 individuals in the autumn of 2023 showed significant level of total S1-RBD antibodies with a correlation of R = 0.70. However, levels in DSS were generally 10 to 100-fold lower than in DBS. Anti-S1-RBD IgG and IgA in DSS demonstrated a correlation of R = 0.6.
Keywords: Antibody isotypes; Antibody proximity extension assay; Dried blood spot (DBS); Dried saliva spot (DSS); IgA; IgG; IgM; Immunoassays; Real-time PCR; SARS-CoV-2 antibody; Vaccination.