Tan E, Abrams-Ogg A C G, Defarges A, Bienzle D
Department of Pathobiology, University of Guelph, 50 Stone Road East, Guelph, Ontario N1G 2W1, Canada.
Department of Clinical Studies, University of Guelph, 50 Stone Road East, Guelph, Ontario N1G 2W1, Canada.
J Comp Pathol. 2014 Jul;151(1):67-79. doi: 10.1016/j.jcpa.2014.02.005. Epub 2014 Feb 26.
Automated analysis of bone marrow (BM) aspirates is a useful 'pre-microscopical' screen to identify hypocellular samples and those with potentially abnormal cells. In order to determine whether automated analysis could also be used to identify haemopoietic abnormalities, EDTA-anticoagulated BM aspirates from 43 dogs were analysed using the Advia 2120 instrument. Corresponding Wright-stained BM smears were evaluated microscopically to determine smear quality, cell composition and 500-cell differential counts, and correlation to automated analysis parameters was computed. Leucocyte cytograms generated by the automated analyzer were scrutinized and compared with those of 'normal' BM. Twenty-three neoplastic and 20 non-neoplastic samples were analysed, including samples from 10 cases of acute myeloid leukaemia, four cases of acute lymphocytic leukaemia, four cases of chronic lymphocytic leukaemia, one case of chronic neutrophilic leukaemia, three cases of multiple myeloma, one case of myelodysplastic syndrome, five cases of non-regenerative immune-mediated haemolytic anaemia, one case of immune-mediated neutropenia, three cases of immune-mediated thrombocytopenia, six cases of inflammatory disease, three samples with myelotoxicity and two samples analysed for staging of neoplasia. Automated white blood cell (WBC) counts correlated significantly with smear cellularity, particle cellularity and particle number. There was a significant difference in WBC counts of samples with insufficient versus sufficient particles. Significant correlations between Advia percent neutrophils and microscopical determination of marrow segmented neutrophils/neutrophilic granulocyte reserve, Advia percent lymphocytes and microscopical determination of lymphocytes/rubricytes, Advia percent large unstained cells and microscopical determination of myeloblasts/promyelocytes and between Advia percent eosinophils and manual determination of eosinophils were identified. This suggested that Advia WBC counts may be used to approximate BM sample quality and that Advia differential counts may predict marrow granulocyte reserve and lymphocyte/rubricyte stores. Distinct and consistent alterations in cytogram patterns were observed in cases of acute leukaemia, but were less obvious in chronic leukaemia. Complete automated BM analysis was performed in approximately 2 min, while staining and coverslipping of BM slides required approximately 30 min. Hence, although automated analysis should not supplant microscopical evaluation of BM, it can provide useful ancillary information in a short time and flag potentially inadequate or abnormal samples.
骨髓（BM）穿刺液的自动化分析是一种有用的“显微镜前”筛查方法，可用于识别细胞减少的样本以及含有潜在异常细胞的样本。为了确定自动化分析是否也可用于识别造血异常，使用Advia 2120仪器对43只犬的乙二胺四乙酸（EDTA）抗凝骨髓穿刺液进行了分析。对相应的瑞氏染色骨髓涂片进行显微镜评估，以确定涂片质量、细胞组成和500个细胞的分类计数，并计算与自动化分析参数的相关性。仔细检查自动分析仪生成的白细胞细胞图，并与“正常”骨髓的细胞图进行比较。分析了23份肿瘤性和20份非肿瘤性样本，包括来自10例急性髓系白血病、4例急性淋巴细胞白血病、4例慢性淋巴细胞白血病、1例慢性中性粒细胞白血病、3例多发性骨髓瘤、1例骨髓增生异常综合征、5例非再生性免疫介导的溶血性贫血、1例免疫介导的中性粒细胞减少症、3例免疫介导的血小板减少症、6例炎症性疾病、3份有骨髓毒性的样本以及2份用于肿瘤分期分析的样本。自动白细胞（WBC）计数与涂片细胞密度、颗粒细胞密度和颗粒数量显著相关。颗粒不足与充足的样本之间的WBC计数存在显著差异。确定了Advia中性粒细胞百分比与骨髓分叶核中性粒细胞/中性粒细胞储备的显微镜测定、Advia淋巴细胞百分比与淋巴细胞/幼红细胞的显微镜测定、Advia大未染色细胞百分比与原始粒细胞/早幼粒细胞的显微镜测定以及Advia嗜酸性粒细胞百分比与嗜酸性粒细胞手工测定之间的显著相关性。这表明Advia WBC计数可用于评估骨髓样本质量，Advia分类计数可预测骨髓粒细胞储备和淋巴细胞/幼红细胞储存。在急性白血病病例中观察到细胞图模式有明显且一致的改变，但在慢性白血病中不太明显。完整的骨髓自动分析大约需要2分钟，而骨髓涂片的染色和封片大约需要30分钟。因此，虽然自动化分析不应取代骨髓的显微镜评估，但它可以在短时间内提供有用的辅助信息，并标记潜在不足或异常的样本。
