直肠黏膜活检组织行钙视网膜蛋白、S100及PGP9．5联合染色对先天性巨结肠症的诊断价值

Calretinin,S100 and protein gene product 9. 5 immunostaining of rectal suction biopsies in the diagnosis of Hirschsprung＇disease

目的 评估直肠黏膜负压吸引活检组织行钙视网膜蛋白（CR）、S100蛋白（S100）及蛋白基因产物9.5 （PGP9.5）染色诊断先天性巨结肠症（HD）的准确性.方法 2014年6月至2017年1月,华中科技大学同济医学院附属协和医院就诊的疑似HD患儿318例,在进行详细的病史调查及体检后行直肠黏膜负压吸引活检,取得的标本进行CR、S100、PGP9.5免疫组织化学染色,观察是否有神经节细胞及粗大增生的神经纤维来诊断HD,并与术后病理检查结果比较,评估其诊断HD的准确性.结果 318例患者进行该诊断方法,其中219（69％）例显示有神经节细胞,99例（31％）缺乏神经节细胞,在这99例中,S-100蛋白及PGP9.5染色分别显示有92％和93％的患儿含有黏膜下粗大的神经纤维.132例患者最终行结肠切除术,其中,99例术后病检证实了HD.该方法的准确性和特异性分别为97.0％ [95％可信区间（CI）：88.0％ ～99.0％]和100.0％ （95％ CI：97.0％～100.0％）,阳性预测值为94.3％ （95％ CI：87.0％ ～ 99.0％）,阴性预测值为99.1％ （95％ CI：95.0％～100.0％）.结论 CR、S-100和PGP9.5染色之间能通过优势互补,提高直肠黏膜负压吸引活检诊断HD的精准性.

Objective To evaluate the sensitivity and specificity of calretinin （CR） combinedwith S100 proteins （S100） and protein gene product 9.5 （PGP9.5） immunostaining of rectal suction biopsies for the diagnosis of Hirschsprung＇s disease （HD）.Methods We conducted a prospective study and collected 318 patients with suspicious HD in Wuhan Union Hospital.All children included underwent the rectal suction biopsy after careful history and physical examinations to confirm or exclude the HD.We applied a protocol using calretinin combined with S100 and PGP9.5 immunostaining for the identification of ganglion cells and hypertrophic nerve trunks.Finally,we evaluated the usefulness of this method for diagnosis of HD.Results Between February 2014 to January 2017,a total of 318 patients were enrolled in our study.Among them,219 patients （69%） had ganglion cells on the initial diagnostic protocol,and 99 cases were devoid of ganglion cells.92% and 93% showed submucosal neural hypertrophy on S100 staining and PGP9.5 staining,respectively.132 patients underwent a colonic resection,and 99 had confirmed aganglionic segment.The sensitivity and speciflcity rates of our diagnostic protocol were 97.0% （95% CI：88.0%-99.0%） and 100.0% （95% CI：97.0%-100.0%） respectively,with positive predictive value and negative predictive value of 94.3% （95% CI：87.0%-99.0%） and 99.1% （95% CI：95.0%-100.0%）,respectively.Conclusion Calretinin coupled with S100 and PGP9.5 immunostaining on suction rectal biopsies is sensitive and specific for diagnosing HD.Each of them offers speciflc advantages,and these complimentary aspects could contribute to an accurate diagnosis.