Uso de sangre total con EDTA y heparina para determinación de fragilidad osmótica y estabilidad a diferentes tiempos de almacenamiento
Fecha
2016
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Universidad de Guayaquil. Facultad de Ciencias Químicas
Resumen
La Fragilidad Osmótica para el diagnóstico de Esferocitosis Hereditaria, es una prueba operador dependiente y de fácil ejecución, no se encuentra disponible en los laboratorios de rutina o de baja complejidad, por lo que es habitualmente referida a Laboratorios de mayor complejidad, sin que exista evidencia local sobre el tipo de muestra recomendada y su estabilidad.
Se realizó un estudio Observacional para establecer diferencias en los resultados de la cuantificación de fragilidad osmótica empleando sangre total EDTA y heparinizada usando técnica estándar de fragilidad osmótica (Parpart) y la estabilidad al almacenamiento.
Se estudiaron 20 muestras de pacientes sanos, empleando sangre total EDTA y sangre total heparinizada, analizadas a la primera hora, 24 y 72 horas. La edad promedio de los sujetos fue de 32. 5 ± 7.8 años, con hemoglobina, hematocrito y VCM promedio de 15.6 ±
1.5 g/dL, 45.6 ± 3.5 %y 87.4 ± 5.6 fL respectivamente.
Cada muestra se analizó en 16 diluciones consecutivas con agua destilada y solución salina, medidas en su absorbancia a 450 nm y relativizadas frente a la absorbancia más alta de las 16 diluciones.
No se evidenciaron diferencias significativas en la cuantificación de fragilidad osmótica independientemente del tipo de muestra. La hemólisis se presentó inicialmente en NaCl,
0.60%, con comportamiento estable hasta las 72 horas refrigeradas.
El 95% de las muestras presentaron algún grado de hemólisis a partir de la solución NaCl
0.52%, independientemente del tipo de muestra. Se pueden transferir los valores de referencia establecidos en otras poblaciones (inicio de hemólisis entre solución 0.5% y 0.3%).
The Osmotic Fragility for diagnosing hereditary spherocytosis, is operator dependent test and not as easy execution, it is not available in standard laboratories, so is commonly referred to laboratories more complex, with no local evidence on recommended type of sample and its stability. An experimental study was conducted to establish differences in the results of the quantification of osmotic fragility using EDTA and heparinized whole blood using standard technique osmotic fragility (Parpart) and storage stability. 20 samples of healthy subjects were studied using EDTA whole blood and heparinized whole blood, analyzed the first hour, 24 and 72 hours. The average age of the subjects was 32. 5 ± 7.8 years, hemoglobin, hematocrit and MCV average of 15.6 ± 1.5 g / dL, 45.6 ± 3.5% and 87.4 ± 5.6 fL respectively. Each sample was analyzed in 16 consecutive dilutions with distilled water and saline, measures its absorbance at 450 nm and relativised facing the highest absorbance of 16 dilutions. No significant differences were found in the quantification of osmotic fragility regardless of sample type. Hemolysis was initially presented in NaCl, 0.60%, with stable performance up to 72 hours refrigerated. 95% of the samples showed some degree of hemolysis from 0.52% NaCl solution, regardless of sample type. You can transfer the benchmarks set in other populations (start of hemolysis solution between 0.5% and 0.3%).
The Osmotic Fragility for diagnosing hereditary spherocytosis, is operator dependent test and not as easy execution, it is not available in standard laboratories, so is commonly referred to laboratories more complex, with no local evidence on recommended type of sample and its stability. An experimental study was conducted to establish differences in the results of the quantification of osmotic fragility using EDTA and heparinized whole blood using standard technique osmotic fragility (Parpart) and storage stability. 20 samples of healthy subjects were studied using EDTA whole blood and heparinized whole blood, analyzed the first hour, 24 and 72 hours. The average age of the subjects was 32. 5 ± 7.8 years, hemoglobin, hematocrit and MCV average of 15.6 ± 1.5 g / dL, 45.6 ± 3.5% and 87.4 ± 5.6 fL respectively. Each sample was analyzed in 16 consecutive dilutions with distilled water and saline, measures its absorbance at 450 nm and relativised facing the highest absorbance of 16 dilutions. No significant differences were found in the quantification of osmotic fragility regardless of sample type. Hemolysis was initially presented in NaCl, 0.60%, with stable performance up to 72 hours refrigerated. 95% of the samples showed some degree of hemolysis from 0.52% NaCl solution, regardless of sample type. You can transfer the benchmarks set in other populations (start of hemolysis solution between 0.5% and 0.3%).
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FRAGILIDAD OSMÓTICA, ESTABILIDAD, EDTA, HEPARINA, HEMÓLISIS