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KMID : 0365819680080020147
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Journal of Pusan Medical College
1968 Volume.8 No. 2 p.147 ~ p.152
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Peroxidase Reaction of Leukocytes in Various Diseases
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Abstract
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Reagents
(1) Ninety-five per cent methanol. Absolute methanol is diluted by the addition of distilled water. (2) Phosphate buffer solution, pH 6.4, M/15, and M/150
(3) Mixture of ammonium bromide and phosphate buffer: Equal amounts of one per cent aqueous ammonium bromide solution and phosphate buffer solution, pH 6.4, M/15 are mixed.
(4) Benzidine H2O2: Dissolve 0. 1 gm. of benzidine in 30 ml. of absolute alcohol, then add 70 ml. of distilled water and 2 drops of 3 per cent H2O2.
(5) Diluted Wright´s stain solution; Equal amounts of stock Wright´s stain solution and phosphate buffer solution, pH 6.4, M/150 are mixed.
Procedure
(1) An air-dried blood smear is fixed with 95 per cent methanol for one minute, and the methanol is poured off.
(2) The smear is then covered with diluted Wright´s stain solution for 10 minutes, and washed with tap water.
(3) One ml. of the mixture of ammonium bromide and phosphate buffer is added to the smear, then 2.0 ml. of benzidine H2O2 is added and allow to remain for 3 minutes. (4) The smear is then washed with tap water.
RESULT
Eosinophilic granules give a strongly positive yellowish brown reaction, neutrophilic granules blue black reaction. Lymphocytes and plasmocytes react negatively. The usual cellular structure is well preserved by Wright´s stain.
The intensity of the peroxidase activity is graded as follows.
0, negative reaction
1+, 1-10 granules
2+, 11-30 granules
3+, many, unevenly distributed granules in the cytoplasm
4+, numerous heavy granules filling the cytoplasm
5+, Coalescent granules
Classify 100 neutrophils (band and segmented) and score of the peroxidase activity is calculated. I. The average peroxidase score of neutrophils in normal 24 subjects was 342 ± 46.
2. The average peroxidase score of neutrophils in 14 cases of acute granulocytic leukemia was 272 ± 68, and lower than normal subjects.
3. The average peroxidase score of neutrophils in 15 cases of chronic granulocytic leukemia was 401 ± 39, and higher than normal subjects.
4. The average peroxidase score of neutrophils in 11 cases of lymphocytic leukemia was 260 ± 32, and lower than normal subjects.
5. 53.2 per cent of myeloblasts in 28 cases of acute granulocytic leukemia were peroxidase positive. 6. 46 per cent of myeloblasts in 9 cases of chronic granulocytic leukemia were peroxidase positive.
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KEYWORD
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