Paper-based assay for red blood cell antigen typing by the indirect antiglobulin test

Natasha Yeow, Heather McLiesh, Liyun Guan, Wei Shen, Gil Garnier

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

Abstract

A rapid and simple paper-based elution assay for red blood cell antigen typing by the indirect antiglobulin test (IAT) was established. This allows to type blood using IgG antibodies for the important blood groups in which IgM antibodies do not exist. Red blood cells incubated with IgG anti-D were washed with saline and spotted onto the paper assay pre-treated with anti-IgG. The blood spot was eluted with an elution buffer solution in a chromatography tank. Positive samples were identified by the agglutinated and fixed red blood cells on the original spotting area, while red blood cells from negative samples completely eluted away from the spot of origin. Optimum concentrations for both anti-IgG and anti-D were identified to eliminate the washing step after the incubation phase. Based on the no-washing procedure, the critical variables were investigated to establish the optimal conditions for the paper-based assay. Two hundred ten donor blood samples were tested in optimal conditions for the paper test with anti-D and anti-Kell. Positive and negative samples were clearly distinguished. This assay opens up new applications of the IAT on paper including antibody detection and blood donor-recipient crossmatching and extends its uses into non-blood typing applications with IgG antibody-based diagnostics. 

Original languageEnglish
Pages (from-to)5231-5238
Number of pages8
JournalAnalytical and Bioanalytical Chemistry
Volume408
Issue number19
DOIs
Publication statusPublished - 1 Jul 2016

Keywords

  • Antibody
  • Blood typing
  • Coombs test
  • IgG
  • Indirect antiglobulin test
  • Paper diagnostics

Cite this

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abstract = "A rapid and simple paper-based elution assay for red blood cell antigen typing by the indirect antiglobulin test (IAT) was established. This allows to type blood using IgG antibodies for the important blood groups in which IgM antibodies do not exist. Red blood cells incubated with IgG anti-D were washed with saline and spotted onto the paper assay pre-treated with anti-IgG. The blood spot was eluted with an elution buffer solution in a chromatography tank. Positive samples were identified by the agglutinated and fixed red blood cells on the original spotting area, while red blood cells from negative samples completely eluted away from the spot of origin. Optimum concentrations for both anti-IgG and anti-D were identified to eliminate the washing step after the incubation phase. Based on the no-washing procedure, the critical variables were investigated to establish the optimal conditions for the paper-based assay. Two hundred ten donor blood samples were tested in optimal conditions for the paper test with anti-D and anti-Kell. Positive and negative samples were clearly distinguished. This assay opens up new applications of the IAT on paper including antibody detection and blood donor-recipient crossmatching and extends its uses into non-blood typing applications with IgG antibody-based diagnostics. ",
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Paper-based assay for red blood cell antigen typing by the indirect antiglobulin test. / Yeow, Natasha ; McLiesh, Heather; Guan, Liyun; Shen, Wei; Garnier, Gil.

In: Analytical and Bioanalytical Chemistry, Vol. 408, No. 19, 01.07.2016, p. 5231-5238.

Research output: Contribution to journalArticleResearchpeer-review

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