REDQUANT 및 CELLQUANT kit를 이용한 PNH 진단
Diagnosis of Paroxysmal Nocturnal Hemoglobinuria (PNH) by the REDQUANT and the CELLQUANT kits
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배경 : PNH는 조혈모세포의 질환으로서 여러 계열의 세포에세포막 결합 GPI-anchored 단백의 발현 장애를 특징으로 한다.상품화된 REDQUANT 및 CELLQUANT 키트(Biocytex, Mar-seille,France)를 사용해서 PNH 진단에 검토하였다.방법 : 다양한 정도의 말초 혈구 감소증을 동반한 70명의 환자와 건강대조군 16명을 대상으로 하였다. REDQUANT 및 CEL-LQUANT 키트를 사용해서 Coulter Epics XL로 분석하였다. 사용 지침에 따라 CD55 및 CD59 항원 결핍 분획이 3% 이상일 때양성으로 하였고, 네 가지 항원 중 세 개 항원 이상이 양성이면PNH로 진단하였다.결과 : CD55/CD59 결핍 적혈구/과립구 분획의 평균은 환자군과 대조군에서 각 0.3, 3.1, 3.5, 10.0, 그리고 0.1, 1.0, 0.3, 9.7이었다. 70명의 환자 중 4항원 및 3항원이 결핍되어 PNH로 진단가능했던 예가 각각 2예, 1예씩 있었고, 두 항원이 감소된 예도 2예 있었다. PNH라고 할 수는 없으나 CD59 결핍 과립구 분획만3% 이상으로 증가된 경우도 환자군 중 30예, 대조군 중 1예에서관찰되었다. CD59 결핍 적혈구도 환자군 및 대조군에서 각각 6예와 2예 있었고, CD55 결핍 과립구도 환자군 1예에서 관찰되었다. 결론 : REDQUANT/CELLQUANT 키트는 방법이 표준화되어 있고 대조군을 필요로 하지 않는 점에서 장점이 있으나 PNH가 아니면서 특히 CD59 결핍 과립구 분획이 증가되는 경우가 있어서 판독에 주의가 요구된다.
Abstract
Background : Paroximal nocturnal hemoglobinuria (PNH) is a disorder of the pluripotent stem cells resulting in a deficient expression of membrane-bound GPI-anchored proteins in different cell types. We evaluated REDQUANT and CELLQUANT kits (Biocytex, Marseille, France) for PNH test. Methods : Seventy patients with peripheral blood cytopenia and 16 healthy controls were studied. RBCs and granulocytes were tested for CD55 and CD59 expression using the REDQUANT and CELLQUANT kits and an Epics XL flow cytometer. According to the manufacturer’s instruction, results were interpreted abnormal when more than 3% of cells were deficient in the expression of CD55 or CD59, and a test was considered positive for PNH if three of the four markers tested were abnormal. Results : The percentage of CD55/CD59 deficient RBCs and granulocytes was 0.3/3.1 and 3.5/ 10.0, respectively, in the patient group, and 0.1/1.0 and 0.3/9.7, respectively, in the control group. PNH was diagnosed in three patients who had a deficiency in the expression of three or four antigens; two other patients showed a deficiency in two antigens. There were many who had CD59 deficiency only: on granulocytes in 30 patients and 11 controls, and on RBCs in 6 patients and 2 controls. One patient had CD55 deficient granulocytes. Conclusions : The REDQUANT/CELLQUANT kit is a standardized method and does not require normal samples as the control, but one should be cautious in interpreting the results showing CD59 expression on granulocytes
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