Reliability of 400-cell and 5-field leukocyte differential counts for equine bronchoalveolar lavage fluid
Reliability of 400-cell and 5-field leukocyte differential counts for equine bronchoalveolar lavage fluid
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DOI：
10.1111/vcp.12013
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发表时间：
2013-03-01
影响因子：
1.2
通讯作者：
Leguillette, Renaud
中科院分区：
文献类型：
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作者：
Fernandez, Nicole J.;Hecker, Kent G.;Leguillette, Renaud
背景 对马支气管肺泡灌洗 (BAL) 液中肥大细胞和嗜酸性粒细胞的可靠计数非常重要，因为这些细胞百分比的小幅增加支持炎症性气道疾病 (IAD) 的临床诊断。 IAD 也会导致 BAL 中性粒细胞增加，但由于 IAD 和复发性气道阻塞 (RAO) 之间存在重叠，因此不具有特异性。目的 本研究的目的是评估标准 400 细胞白细胞分类计数的可靠性，以及评估马 BAL 液体细胞离心制剂中 500 倍放大倍率下的 5 个显微视野的替代方法。方法采用 400 细胞和 5 视野白细胞分类计数方法对 60 匹有或没有肺部炎症的马的 BAL 样本进行评估。通过计算和比较组内相关系数（ICC）来评估每种白细胞类型计数的可靠性。通过比较具有不同细胞密度的载玻片的ICC，进一步评估肥大细胞计数的可靠性。结果 5 视野法对所有细胞类型的可靠性均较高；然而，总体而言，方法之间的差异并不具有统计学意义。两种方法的中性粒细胞可靠性都很高（ICC > 0.90）。只有在细胞密度较高的载玻片上采用 5 视野方法才能实现肥大细胞的足够可靠性 (ICC > 0.85)。结论 当使用标准的400个细胞分类计数方法时，肥大细胞的计数不可靠，而在具有较高细胞密度的载玻片上的5视野方法达到了可接受的重现性。两种方法的中性粒细胞百分比均高度可靠。
Background Reliable enumeration of mast cells and eosinophils in equine bronchoalveolar lavage (BAL) fluid is important because small increases in the percentages of these cells support the clinical diagnosis of inflammatory airway disease (IAD). Increases in BAL neutrophils also occur with IAD but are not specific due to overlap between IAD and recurrent airway obstruction (RAO). Objectives The objectives of this study were to evaluate the reliability of a standard 400-cell leukocyte differential count and an alternate method evaluating 5 microscopic fields at 500x magnification in equine BAL fluid cytocentrifuged preparations. Methods BAL samples from 60 horses with and without pulmonary inflammation were evaluated using 400-cell and 5-field leukocyte differential counting methods. Reliability of enumeration of each leukocyte type was assessed by calculating and comparing intraclass correlation coefficients (ICC). Reliability of mast cell enumeration was further evaluated by comparing ICCs of slides with different cell densities. Results Reliability was higher for all cell types with the 5-field method; however, overall the difference between methods was not statistically significant. Neutrophil reliability was high (ICC > 0.90) with both methods. Adequate reliability (ICC > 0.85) for mast cells was achieved only with the 5-field method on slides with higher cell density. Conclusion Enumeration of mast cells is unreliable when the standard 400-cell differential counting method is used, whereas the 5-field method on slides with higher cell density reached acceptable reproducibility. Neutrophil percentages were highly reliable with both methods.