Calretinin, S100 and protein gene product 9.5 immunostaining of rectal suction biopsies in the diagnosis of Hirschsprung’ disease

Abstract

Objective To evaluate the sensitivity and specificity of calretinin (CR) combined with 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. Key words: Calretinin; S100 protein; Protein gene product 9.5; Hirschsprung’s disease