Olin J Tod, Burns Kim, Carson Johnny L, Metjian Hilda, Atkinson Jeffrey J, Davis Stephanie D, Dell Sharon D, Ferkol Thomas W, Milla Carlos E, Olivier Kenneth N, Rosenfeld Margaret, Baker Brock, Leigh Margaret W, Knowles Michael R, Sagel Scott D
Department of Pediatrics, The Children's Hospital and University of Colorado School of Medicine, Aurora, Colorado.
Pediatr Pulmonol. 2011 May;46(5):483-8. doi: 10.1002/ppul.21402. Epub 2011 Jan 31.
Examination of ciliary ultrastructure remains the cornerstone diagnostic test for primary ciliary dyskinesia (PCD), a disease of abnormal ciliary structure and/or function. Obtaining a biopsy with sufficient interpretable cilia and producing quality transmission electron micrographs (TEM) is challenging. Methods for processing tissues for optimal preservation of axonemal structures are not standardized. This study describes our experience using a standard operating procedure (SOP) for collecting nasal scrape biopsies and processing TEMs in a centralized laboratory. We enrolled patients with suspected PCD at research sites of the Genetic Disorders of Mucociliary Clearance Consortium. Biopsies were performed according to a SOP whereby curettes were used to scrape the inferior surface of the inferior turbinate, with samples placed in fixative. Specimens were shipped to a central laboratory where TEMs were prepared and blindly reviewed. Four hundred forty-eight specimens were obtained from 107 young children (0-5 years), 189 older children (5-18 years), and 152 adults (> 18 years), and 88% were adequate for formal interpretation. The proportion of adequate specimens was higher in adults than in children. Fifty percent of the adequate TEMs showed normal ciliary ultrastructure, 39% showed hallmark ultrastructural changes of PCD, and 11% had indeterminate findings. Among specimens without clearly normal ultrastructure, 72% had defects of the outer and/or inner dynein arms (IDA), while 7% had central apparatus defects with or without IDA defects. In summary, nasal scrape biopsies can be performed in the outpatient setting and yield interpretable samples, when performed by individuals with adequate training and experience according to an SOP.
对纤毛超微结构的检查仍然是原发性纤毛运动障碍(PCD)的关键诊断测试,PCD是一种纤毛结构和/或功能异常的疾病。获取具有足够可解释纤毛的活检样本并制作高质量的透射电子显微镜图像(TEM)具有挑战性。用于最佳保存轴丝结构的组织处理方法尚未标准化。本研究描述了我们在集中实验室中使用标准操作程序(SOP)收集鼻刮活检样本并处理TEM的经验。我们在黏液纤毛清除遗传疾病联盟的研究地点招募了疑似PCD的患者。活检按照SOP进行,即用刮匙刮取下鼻甲的下表面,样本置于固定剂中。标本被送往中央实验室,在那里制备TEM并进行盲法评估。从107名幼儿(0 - 5岁)、189名大龄儿童(5 - 18岁)和152名成人(>18岁)中获取了448份标本,其中88%的标本适合进行正式解读。成人中合适标本的比例高于儿童。50%的合格TEM显示纤毛超微结构正常,39%显示PCD的标志性超微结构改变,11%的结果不确定。在超微结构未明确正常的标本中,72%存在外动力蛋白臂和/或内动力蛋白臂(IDA)缺陷,而7%存在中央微管结构缺陷,伴或不伴有IDA缺陷。总之,鼻刮活检可以在门诊进行,并且当由经过充分培训和有经验的人员按照SOP进行操作时,可以获得可解读的样本。