A multicenter trial of the effectiveness of zeta-globin enzyme-linked immunosorbent assay and hemoglobin H inclusion body screening for the detection of alpha0-thalassemia trait.

Routine laboratories use a hemoglobin H (HbH) screen to detect alpha-thalassemia carriers of fatal hemoglobin Bart's hydrops fetalis. This test is laborious and has sensitivity concerns. A commercial zeta-globin enzyme-linked immunosorbent assay (ELISA) is effective in detecting Southeast Asian (SEA) alpha-thalassemia. We present results of a study of the effectiveness of carrier detection of ELISA and a shortened HbH screen compared with gap polymerase chain reaction. ELISA was superior to the HbH screen for the SEA alpha0-thalassemia trait. The ELISA and H screen were equal for detection of all carriers encountered and combined were more effective than either test alone. A positive zeta-globin ELISA result is diagnostic of SEA alpha-thalassemia, and routine use of the zeta-globin ELISA in combination with a shortened HbH screen will improve the efficacy of prenatal screening for carriers of hemoglobin Bart's hydrops fetalis through improved detection and referral for follow-up DNA testing.

An improved method for detecting red cells with hemoglobin H inclusions that does not require glass capillary tubes.

alpha-Thalassemia trait is the most common inherited abnormality worldwide. Diagnosis of alpha-thalassemia trait can be difficult as there are no abnormalities detectable by hemoglobin electrophoresis or high-performance liquid chromatography. Detection of individuals with alpha-thalassemia trait, particularly the type present in many Asian populations, is important for genetic counseling purposes, because these individuals are at risk for having offspring with hemoglobin Bart's hydrops fetalis, a fatal condition. The best routine diagnostic method to detect individuals with alpha-thalassemia trait is staining reticulocyte-enriched red cell preparations with brilliant cresyl blue to detect hemoglobin H inclusions. Current methods use centrifugation of microhematocrit tubes to enrich for reticulocytes, which presents a laboratory safety hazard. In this report, we describe an alternative technique to enrich for reticulocytes that does not require glass capillary tubes, but is as effective as the capillary tube method for reticulocyte enrichment and detection of cells containing hemoglobin H inclusions.

Hypofunción esplénica espontánea en lupus eritematoso sistémico y sindrome de Sjögren primario / Spontaneous splenic hypofunction in systemic lupus erythematosus and primary Sjogren syndrome

El objetivo del presente trabajo fue evaluar la existencia de hipoesplenismo funcional en enfermedades autoinmunes. La hipofunción esplénica se determinó por la presencia de cuerpos de Howell-Jolly en eritrocitos de la sangre y posterior centellografía esplénica con radiocoloides marcados con 99Tc, en pacientes en los que se habían descartado otras posibles causas de cuerpo de Howell-Jolly. Se estudiaron extendidos de sangre periférica de 174 pacientes con enfermedades autoinmunes y de 32 otras enfermedades de patogenia inmunológica y 63 indivíduos normales. Se encontró evidencia de hipoesplenia funcional en 4 de 79 pacientes con Lupus Eritematoso Sistémico (LES) y en 2 de 18 casos de Síndrome de Sjorgren Primario (SSP). En uno de los pacientes de LES con hipoesplenia funcional y leve esplenomegalia, se demostró ausencia de cuerpos Howell-Jolly en un estudio realizado 11 meses después de la primera evaluación. En el grupo control, un paciente con leucemia linfática crónica (LLC) y esplenomegalia tenía evidencia de hipoesplenismo. Los resultados obtenidos revelan: 1) Las enfermedades autoinmunes que con mayor frecuencia presentan hipoesplenia funcional son el LES y SSP; 2) La hipoesplenia funcional en enfermedades autoinmunes puede ser transitoria. 3) Esplenomegalia e imagen centelleográfica aumentada del bazo no descartan hipoesplenia

Hypofunción esplénica espontánea en lupus eritematoso sistémico y sindrome de Sj÷gren primario / Spontaneous splenic hypofunction in systemic lupus erythematosus and primary Sjogren syndrome

El objetivo del presente trabajo fue evaluar la existencia de hipoesplenismo funcional en enfermedades autoinmunes. La hipofunción esplénica se determinó por la presencia de cuerpos de Howell-Jolly en eritrocitos de la sangre y posterior centellografía esplénica con radiocoloides marcados con 99Tc, en pacientes en los que se habían descartado otras posibles causas de cuerpo de Howell-Jolly. Se estudiaron extendidos de sangre periférica de 174 pacientes con enfermedades autoinmunes y de 32 otras enfermedades de patogenia inmunológica y 63 indivíduos normales. Se encontró evidencia de hipoesplenia funcional en 4 de 79 pacientes con Lupus Eritematoso Sistémico (LES) y en 2 de 18 casos de Síndrome de Sjorgren Primario (SSP). En uno de los pacientes de LES con hipoesplenia funcional y leve esplenomegalia, se demostró ausencia de cuerpos Howell-Jolly en un estudio realizado 11 meses después de la primera evaluación. En el grupo control, un paciente con leucemia linfática crónica (LLC) y esplenomegalia tenía evidencia de hipoesplenismo. Los resultados obtenidos revelan: 1) Las enfermedades autoinmunes que con mayor frecuencia presentan hipoesplenia funcional son el LES y SSP; 2) La hipoesplenia funcional en enfermedades autoinmunes puede ser transitoria. 3) Esplenomegalia e imagen centelleográfica aumentada del bazo no descartan hipoesplenia (AU)

[Spontaneous splenic hypofunction in systemic lupus erythematosus and primary Sjögren syndrome]. / Hipofunción esplénica espontánea en lupus eritematoso sistémico y síndrome de Sjogren primario.

UNLABELLED: The objective of this study was to evaluate hyposplenism in autoimmune diseases by the presence of Howell-Jolly bodies in blood erythrocytes and 99Tc spleen scan. Blood smears of 174 patients with autoimmune diseases and 126 controls were studied. Other possible causes for the presence of Howell-Jolly bodies were excluded. Evidence of hyposplenism was demonstrated in 4 of 79 patients with Systemic Lupus Erythematosus (SLE) and in 2 of 18 cases of primary Sjögren Syndrome (PSS), whereas no hyposplenism was found in the remaining cases of other autoimmune diseases. In one of the patients with SLE, hyposplenism was transient. Among the control cases, a patient with Chronic Lymphocytic Leukemia with splenomegaly presented hyposplenism. IN CONCLUSION: 1) Hyposplenism is more frequently found in SLE and PSS than in other autoimmune diseases. 2) Hyposplenism in autoimmune diseases can be transient. 3) Splenomegaly and enlarged spleen scan do not exclude hyposplenism.

Efficacy of a modified improved technique for detecting red cell haemoglobin H inclusions.

Alpha-thalassaemia is a common disease in Taiwan. A feature useful in diagnosis is the excess of of beta-chains that result from impaired alpha-chain production. These excess chains assemble into beta 4 tetramers (i.e. Hb H) which can be detected by its rapid anodal migration on alkaline electrophoretic media as seen in Hb H disease. However, this technique cannot, and conventional Hb H inclusion staining rarely can visualize alpha-thalassaemic traits due to small quantities of Hb H formed in these patients. The staining for Hb H inclusion bodies uses brilliant cresyl blue (BCB) or methylene blue (MB) as an oxidant to denature Hb H as intracellular inclusions. We have improved the technique of Jones (who modified the original method in order to obtain enriched young red cells) by prolonging the incubation time from 30 min to 3 h. The sensitivity of this modified improved method was 91% for detecting obligatory alpha-thalassaemic traits as shown in table. No false positive results were seen in beta-thalassaemia or in others. We believe this can be used as a confirmatory test in heterozygous alpha 1-thalassaemia and homozygous alpha 2-thalassaemia.