Efficiency of Rapid Brilliant Cresyl Blue Staining for Detecting Hemoglobin H Inclusion Bodies in Red Blood Cells

Abstract

Hemoglobin H disease can be diagnosed by the identification of hemoglobin (Hb) H inclusion in red blood cells; however, a long incubation period of the blood-dye mixture is required for the testing procedure. The objective of this study was to investigate the impact of shortening the Brilliant cresyl blue staining duration on the ability to identify HbH inclusion bodies in red blood cells. Blood samples were collected from 28 HbH disease patients and 30 healthy individuals. Hemoglobin typing was conducted by using HPLC. Incubation of the blood with Brilliant cresyl blue at 37^oC for 30 and 120 minutes were set up for the detection of HbH inclusion bodies. Paired student’s t-test were utilized to assess the reliability of the test results and the variances in the number of HbH inclusion bodies. There were no HbH inclusion bodies found in the healthy control bloods. Conversely, the HbH inclusion bodies were detected in all HbH disease samples at the incubation times of 30 and 120 minutes. The quantities of HbH inclusion bodies at 30- and 120-minute incubation times were 43.7 ± 23.6 and 39.9 ± 22.1, respectively. There was no statistical significance in both incubation times (p < 0.05). These reveal significant amounts of HbH inclusion bodies detected following 30 minutes staining. Therefore, the identification of HbH inclusion elements using this 30 minutes staining method could facilitate a better turnaround time.