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Identification of RHD allelic variants discovered by atypical typing results on the NEO/Echo platforms
J P Hudgins1, C Matsushita2, C W Tuma3
1Department of Pathology, Keck Hospital of USC and USC Norris Cancer Hospital, and Medical Director, Transfusion Medicine Service, LAC+USC Medical Center, Department of Pathology , Suite A7, 1200 N. State Street , Los Angeles, CA 90033.
Abstract:
Some individuals are at risk of anti-D alloimmunization if they inherit D antigens that are qualitatively and/or quantitatively different than wild-type D. We hypothesized that patients who showed serologically inconsistent, weak, or historically discordant D typing results by microplate direct agglutination (MDA) on NEO or Echo (Immucor, Norcross, GA) might be at risk of carrying RHD allelic variants. The present study was designed to evaluate patients with RHD allelic variants if they presented with weakly reactive D typing results on the NEO or Echo. Patients were selected for RHD genotyping if their specimens showed weak reactivity with either series 4 or series 5 anti-D typing reagent, if the strength of reactivity was ≤1+ on the NEO or Echo, or if historical or current D typing results were discordant with current results. Patients selected for RHD genotyping were also tested by saline tube testing using the same anti-D series 4 and 5 reagents. Genotyping was performed by the Immucor genotyping laboratory in Warren, NJ. Of 80 patients whose samples met study inclusion, 52 (65.0%) were found to have RHD allelic variants. Sixteen patients (20.0%) expressed possible Ceppellini effect reactivity. Most importantly, 51.25 percent of the patients who presented with weakly reactive D typing results by MDA testing on the NEO (≤1+) or Echo (≤1+) had RHD allelic variants that were associated with the potential for anti-D alloimmunization. Laboratories that use MDA testing on the Neo or Echo for D typing should consider that female patients of childbearing age might be at risk of anti-D alloimmunization if they are classified as D+ based on weakly reactive D typing results.
Abstract:
Some individuals are at risk of anti-D alloimmunization if they inherit D antigens that are qualitatively and/or quantitatively different than wild-type D. We hypothesized that patients who showed serologically inconsistent, weak, or historically discordant D typing results by microplate direct agglutination (MDA) on NEO or Echo (Immucor, Norcross, GA) might be at risk of carrying RHD allelic variants. The present study was designed to evaluate patients with RHD allelic variants if they presented with weakly reactive D typing results on the NEO or Echo. Patients were selected for RHD genotyping if their specimens showed weak reactivity with either series 4 or series 5 anti-D typing reagent, if the strength of reactivity was ≤1+ on the NEO or Echo, or if historical or current D typing results were discordant with current results. Patients selected for RHD genotyping were also tested by saline tube testing using the same anti-D series 4 and 5 reagents. Genotyping was performed by the Immucor genotyping laboratory in Warren, NJ. Of 80 patients whose samples met study inclusion, 52 (65.0%) were found to have RHD allelic variants. Sixteen patients (20.0%) expressed possible Ceppellini effect reactivity. Most importantly, 51.25 percent of the patients who presented with weakly reactive D typing results by MDA testing on the NEO (≤1+) or Echo (≤1+) had RHD allelic variants that were associated with the potential for anti-D alloimmunization. Laboratories that use MDA testing on the Neo or Echo for D typing should consider that female patients of childbearing age might be at risk of anti-D alloimmunization if they are classified as D+ based on weakly reactive D typing results.
Insights
Weakly reactive D typing results may indicate RHD allelic variants, increasing the risk of anti-D alloimmunization in women of childbearing age. Further RHD genotyping is recommended for accurate risk assessment.
Area of Science:
- Blood banking and transfusion medicine
- Genetics and genomics
- Immunology
Background:
- Individuals with D antigens differing from wild-type are at risk of anti-D alloimmunization.
- Weak or inconsistent D typing results may suggest underlying RHD allelic variants.
Purpose of the Study:
- To evaluate patients with RHD allelic variants presenting with weakly reactive D typing results.
- To identify individuals at risk for anti-D alloimmunization based on D antigen typing.
Main Methods:
- Patients with weak (≤1+) or discordant D typing results on NEO or Echo microplate direct agglutination (MDA) were selected.
- RHD genotyping was performed on selected patient specimens.
- Saline tube testing was used as a comparative method.
Main Results:
- 52 out of 80 (65.0%) patients had RHD allelic variants.
- Weakly reactive D typing results (≤1+) on MDA were associated with RHD allelic variants in 51.25% of cases.
- 16 patients (20.0%) showed Ceppellini effect reactivity.
Conclusions:
- Weakly reactive D typing results on NEO or Echo MDA may indicate RHD allelic variants.
- Female patients of childbearing age classified as D+ with weak reactivity are at risk for anti-D alloimmunization.
- RHD genotyping should be considered for patients with equivocal D typing results.
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