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Differentiation of Sickle Cell Zygosity Utilizing a Sodium Metabisulfite Method
Author(s) -
Le Austin,
Uppugunduri Manoja,
Korducki John,
Randolph Tim
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.636.7
Subject(s) - sickle cell anemia , sickle cell trait , genotype , medicine , hemoglobinopathy , cell , disease , physiology , pediatrics , immunology , pathology , biology , genetics , gene
Hemoglobinopathies are the most prevalent monogenetic disorder worldwide, and sickle cell disease is the most prevalent and severe structural hemoglobinopathy. Globally, over 85% of the sickle cell gene penetrance occurs in sub‐Saharan Africa (64.4%) and Arab‐India (22.7%) which contains the poorest countries on earth. Diagnosing sickle cell disease remains problematic in these populations where poverty severely limits clinic budgets, country infrastructure, and healthcare education making the use of modern diagnostic methods impractical. By modifying an established sodium metabisulfite method to detect sickle cells, we developed a simple, rapid, cost‐effective, microscopic method to distinguish the homozygous genotype (SS) seen in sickle cell disease (SCD) from the heterozygous genotype (AS) seen in sickle cell trait (SCT). We hypothesized that the two zygosities can be differentiated by observing the number, intensity, and rate of sickle cell formation over time. A solution of 2% sodium metabisulfite was prepared and 15 μL was mixed with 15 uL of patient blood and 5uL of the mixture was transferred to a microscope slide for evaluation using 100x oil immersion brightfield microscopy. Sickle cells were counted in one hour intervals for three hours and judged for sickling intensity on a 0–4+ Likert Scale. It was determined that 4+ sickle cells at 3 hours incubation optimally differentiated SS from AS genotypes. SS samples had an average of 135, 4+ sickle cells (N=10) compared to 12, 4+ sickle cells in the sickle cell trait samples (N=5) at three hours. Although more data are being collected, this inexpensive method can potentially be used to diagnose SCD and SCT, benefiting countries where there are limited resources. Support or Funding Information AL is supported by DeNardo Education and Research Foundation Fellowship. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .