Tracy Stokol, Alaina M Kosko, Dorothee Bienzle, Kristina Meichner, Nora L Springer, Michael Byron, Martha Hoffman
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引用次数: 0
Abstract
Background: Assessment of alkaline phosphatase (ALP) activity in cytologic smears is used as a phenotyping tool for hematopoietic and solid tissue neoplasms in dogs. Different procedures that vary in substrate-dye combinations, fixative, incubation times, and nuclear counterstains are available for detection of ALP activity. It is unknown if these procedures provide comparable results for phenotyping tumors in the same animal.
Objective: To compare results obtained with three ALP cytochemical procedures in blasts in blood and tissue aspirate smears from dogs with previously diagnosed leukemia: naphthol-AS-MX phosphate/fast blue (fast blue), 5-bromo-4-chloro-3-indoxyl-phosphate/nitroblue tetrazolium (BCIP/NBT), and naphthol-AS-BI-phosphate/fast red violet LB (fast red).
Methods: Smears of blood, bone marrow, and lymph node from 54 dogs prospectively enrolled in a multi-institutional study on acute leukemia were stained and assessed. One observer counted the percentage of positive blasts and quantified staining intensity on a scale of 1-3. Cases were classified as acute myeloid leukemia, lymphoblastic leukemia/lymphoma, acute leukemia of ambiguous lineage, or acute lineage-negative leukemia based on flow cytometric analysis and myeloperoxidase cytochemical staining.
Results: The fast blue procedure yielded a significantly higher median percentage of ALP-positive blasts (48%) than the BCIP/NBT (46%) or fast red (42%) procedure despite similar median staining intensities. The proportion of samples that would have been classified as ALP-positive was similar (40/54 fast blue, 39/54 BCIP/NBT, and 37/54 fast red).
Conclusions: Our results indicate that the three procedures can be used interchangeably for determining ALP activity in blasts of dogs with leukemia.
背景：细胞学涂片中碱性磷酸酶（ALP）活性的评估被用作犬造血和实体组织肿瘤的表型工具。不同的底物-染料组合、固定液、孵育时间和核反染色方法可用于检测ALP活性。目前尚不清楚这些方法是否能为同一动物的肿瘤表型分析提供可比较的结果。目的：比较三种ALP细胞化学方法对先前诊断为白血病的狗的血液和组织抽吸涂片的结果：萘酚- as - mx磷酸/快蓝（快蓝），5-溴-4-氯-3-吲哚-磷酸/硝基蓝四唑（BCIP/NBT）和萘酚- as - bi -磷酸/快红紫LB（快红）。方法：对54只狗的血液、骨髓和淋巴结涂片进行染色和评估，这些狗被前瞻性地纳入了一项多机构急性白血病研究。一名观察员计数阳性胚的百分比和定量染色强度在1-3级。根据流式细胞术分析和骨髓过氧化物酶细胞化学染色，将病例分为急性髓系白血病、淋巴细胞白血病/淋巴瘤、谱系不明的急性白血病和急性谱系阴性白血病。结果：尽管中位染色强度相似，快速蓝色处理产生的alp阳性细胞中位数百分比（48%）明显高于BCIP/NBT处理（46%）或快速红色处理（42%）。被归类为alp阳性的样本比例相似（40/54快速蓝色，39/54 BCIP/NBT和37/54快速红色）。结论：我们的结果表明，这三种方法可互换用于测定白血病犬原细胞ALP活性。
期刊介绍：
Veterinary Clinical Pathology is the official journal of the American Society for Veterinary Clinical Pathology (ASVCP) and the European Society of Veterinary Clinical Pathology (ESVCP). The journal''s mission is to provide an international forum for communication and discussion of scientific investigations and new developments that advance the art and science of laboratory diagnosis in animals. Veterinary Clinical Pathology welcomes original experimental research and clinical contributions involving domestic, laboratory, avian, and wildlife species in the areas of hematology, hemostasis, immunopathology, clinical chemistry, cytopathology, surgical pathology, toxicology, endocrinology, laboratory and analytical techniques, instrumentation, quality assurance, and clinical pathology education.
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