Application of Flow Cytometry Combined Fluorescence in Situ Hybridization to Indentify the Lymphocyte Subtypies with Epstein-Barr Virus Infection

• Published in: Zhongguo shi yan xue ye xue za zhi Vol. 30; no. 3; p. 897
• Main Authors: Su, Hong-Yu, Shu, Yi, Fu, Guo, Liu, Zi-Yang, Zhu, Dan, Zeng, La-Mei, Ma, De-Yu, Zou, Lin
• Format: Journal Article
• Language: Chinese
• Published: China 01.06.2022
• Subjects: Epstein-Barr Virus Infections; Flow Cytometry - methods; Herpesvirus 4, Human; Humans; In Situ Hybridization, Fluorescence - methods; Lymphocyte Subsets; EBV; differential diagnosis; EBER; Flow-FISH
• ISSN: 1009-2137
• DOI: 10.19746/j.cnki.issn.1009-2137.2022.03.038

To establish the technique that take the advantages of flow cytometry combined fluorescence in situ hybridization (Flow-FISH) to identify the Epstein-Barr virus(EBV) infected lymphocyte subtypies in patients' peripheral blood sample. Peripheral Blood monocyte from 9 patients with EBV infection enrolled at Children's Hospital in Chongqing Medical University were isolated by Ficoll-paque centrifugal separation. The expressions of EBER1, EBER2 in cell were detected by qRT-PCR. The surface markers of cell were detected by Flow cytometry after staining with their antibodies. The cell was treated Fix-Permeabilization Buffer before hybridization with fluorescent labeled probe at 37 ℃ overnight. The cell status, surface markers and targeted mRNA are detected by flow cytometry and fluorescence microscope. It was optimized that the Fix-Permeabilization Buffer and recipe with 0.2% Tween-20 were picked out as providing a good cell integrity and high resolution of surface markers. Hybridization with 20% formamide and 7% d