A comparative study of the gel-centrifugation technique and flow cytometry for the detection of "PNH red cell" populations in the hematological disorders

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A comparative study of the gel-centrifugation technique and flow cytometry for the detection of "PNH red cell" populations in the hematological disorders

 

Author: Man, Kwai-fun
Title: A comparative study of the gel-centrifugation technique and flow cytometry for the detection of "PNH red cell" populations in the hematological disorders
Degree: M.Sc.
Year: 2000
Subject: Blood -- Diseases
Erythrocytes
Hong Kong Polytechnic University -- Dissertations
Department: Multi-disciplinary Studies
Dept. of Nursing and Health Sciences
Pages: ix, 70 leaves : ill. (some col.) ; 30 cm
Language: English
InnoPac Record: http://library.polyu.edu.hk/record=b1541792
URI: http://theses.lib.polyu.edu.hk/handle/200/4481
Abstract: Paroxysmal Nocturnal Hemoglobinuria (PNH) is an acquired clonal disorder due to somatic mutations of the Phosphatidylinositol Glycan A (PIG-A) gene of the X-chromosome that leads to defective biosynthesis of the Glycosylphosphatidylinositol (GPI) anchor. It is characterized by an unusual sensitivity of abnormal red cell populations to complement lysis, due to a complete lack or partial deficiency of complement inhibitory surface GPI-linked protein molecules including CD55 and CD59. PNH has been associated with various hematological disorders. The demonstration of the abnormal PNH red cells can be illustrated by complement mediated lysis test or flow cytometry. In this study, a newly introduced method, the DiaMed-ID Microtyping System for PNH based on Gel-centrifugation Technique was used to detect the presence of CD55 or CD59 defective red cells populations in several hematological disorders including genuine PNH disease. 30 samples with 30 normal controls were analyzed by Sucrose Lysis test and Ham's test to screen for the presence of PNH defective red cells. The first ten samples together with the normal controls were analyzed by the DiaMed-ID Microtyping System to set up the baseline. Another ten samples which were particularly requested for Ham's test, were analyzed by the gel test as well. Due to resource limitation, as a rule, only positive results of gel test would be evaluated by flow cytometry. For the two PNH cases, the gel test results were directly compared with flow cytometric analysis and concordant conclusion was achieved. It was found that a very small population of CD55 or CD59 defective red cells was detected in a few cases by the DiaMed-ID Microtyping System but not by the flow cytometry. Defective red cells were detected in normal subjects as well. Though these findings need further investigations, nevertheless the DiaMed-ID Microtyping System for PNH although non-specific, seems to be an alternative method in screening the abnormal PNH red cell populations in genuine PNH as well as other hematological disorders.

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