Accurate light microscopic diagnosis of South-East Asian ovalocytosis.
Accurate light microscopic diagnosis of South-East Asian ovalocytosis.
复制标题
东南亚卵形细胞增多症的准确光学显微镜诊断。
DOI：
10.1111/ijlh.12900
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发表时间：
2018
影响因子：
3
通讯作者：
KevinBaird,J
中科院分区：
文献类型：
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作者：
Nixon,ChristianP;Satyagraha,AriW;Baird,GraysonL;Harahap,AlidaR;Panggalo,LydiaV;Ekawati,LennyL;Sutanto,Inge;Syafruddin,Din;KevinBaird,J
东南亚卵胞增多症(SAO)是东南亚和美拉尼西亚人群中常见的遗传性红细胞多态，与疟疾流行地区不谋而合。通过对存档的血涂片进行分析，验证光学显微镜作为分子基因分型的诊断替代方法，可以使其前瞻性或回溯性地使用具有成本效益。三名训练有素的显微镜专家，每人评估了同样的971张Giemsa染色的血液薄膜，这些薄膜可以通过聚合酶链式反应确认SAO基因。结果在所评估的红细胞形态参数中，核细胞比卵圆细胞形态更能预测SAO状态(优势比[OR]=19.2；95%可信区间[95%CI]=14.6-25.3；P=0.0001)。以核细胞为中心的显微镜方法的诊断性能依赖于显微镜医生：两位显微镜医生应用这种方法的灵敏度为0.89，特异性为0.93。由于一位显微镜医师将口腔上皮细胞错误地分类为无节细胞(κ=.20)以及金标准和显微镜间(κ=.36)，评分间信度较差，但在排除容易出错的读数后显著提高(分别为κ=.65和=.74)。结论通过无节细胞视觉提示进行SAO的光镜诊断与耗时和昂贵的分子方法相当，但需要特殊的培训，包括成功地区分节细胞和口腔细胞。
IntroductionSoutheast Asian ovalocytosis (SAO) is a common inherited red blood cell polymorphism in Southeast Asian and Melanesian populations, coinciding with areas of malaria endemicity. Validation of light microscopy as a diagnostic alternative to molecular genotyping may allow for its cost-effective use either prospectively or retrospectively by analysis of archived blood smears.MethodsWe assessed light microscopic diagnosis of SAO compared to standard PCR genotyping. Three trained microscopists, each assessed the same 971 Giemsa-stained thin blood films for which SAO genotypic confirmation was available by PCR. Generalized mixed modeling was used to estimate the sensitivity, specificity, positive predictive value, and negative predictive value of light microscopy versus “gold-standard” PCR.ResultsAmong red cell morphologic parameters evaluated, knizocytes, rather than ovalocytic morphology, proved the strongest predictor of SAO status (odds ratio [OR] =19.2; 95% confidence interval [95% CI] = 14.6-25.3; P = <.0001). The diagnostic performance of a knizocyte-centric microscopic approach was microscopist-dependent: two microscopists applied this approach with a sensitivity of 0.89 and a specificity of 0.93. Inter-rater reliability among the microscopists (κ = .20) as well as between gold standard and microscopist (κ = .36) underperformed due to misclassification of stomatocytes as knizocytes by one microscopist, but improved substantially when excluding the error-prone reader (κ = .65 and = .74, respectively).ConclusionLight microscopic diagnosis of SAO by knizocyte visual cue performed comparable to time-consuming and costlier molecular methods, but requires specific training that includes successful differentiation of knizocytes from stomatocytes.