Establishment and Clinical Application of Flow Cytometric Immunobead Array in Detecting Plasma von Willebrand Factor Antigen.
10.7534/j.issn.1009-2137.2020.01.040
- Author:
Bin YAN
1
;
Yang HE
2
;
Shi-Qi LU
3
;
Le QU
4
;
Meng-Qiao XU
2
;
Qi WANG
2
;
Yi-Ming ZHAO
5
;
Chang-Geng RUAN
2
Author Information
1. Department of Clinical Laboratorial Medicine, Nanyang City Cential Hospital, Nanyang 473000, Henan Province, China,The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215006, Jiangsu Province, China.
2. The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215006, Jiangsu Province, China.
3. Department of Emergency, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China.
4. Department of Clinical Laboratorial Medicine, The East District of Suzhou Municipal Hospital, Suzhou 215006, Jiangsu Province, China.
5. The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Suzhou 215006, Jiangsu Province, China,E-mail: zhaoyimingbox@163.com.
- Publication Type:Journal Article
- From:
Journal of Experimental Hematology
2020;28(1):235-241
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish a novel flow cytometric immunobead array (FCIA) for detecting plasma von Willebrand factor antigen (vWF:Ag), and to analyze the clinical value of FCIA in predicting the prognosis of patients with ischemic stroke (IS).
METHODS:Anti-human vWF monoclonal antibody SZ29 IgG was coated on microspheres overnight, the diluted plasma was added after blocking, then incubated with FITC-conjugated sheep-anti-human vWF IgG polyclonal antibody, and finally detected by flow cytometry. The plasma vWF in 21 case of von Willebrand disease (vWD) and 105 controls (CTL) were detected by FCIA and ELISA, so as to carry out methodological assessment. Plasma vWF:Ag of 61 IS patients was detected by FCIA and the data of prognosis followed-up for 2-year were collected.
RESULTS:The linear fitting of FCIA was good (R2=0.99) without significant difference between FCIA and ELISA. The Bland-Altman bias was 1.12% with 95% limits of agreement that spanned from -45.06% to 47.30%, and the slope of the linear regression was 0.97 (r=0.86, P＜0.01). Importantly, the FCIA method was faster than ELISA, and superior to the ELISA in the detection of low levels of vWF:Ag. The levels of vWF:Ag, vWF:GPIbR and vWF:CB in IS patients were significantly higher than those in healthy controls (Z=8.36, 8.71, 6.22, respectively, P＜0.01).
CONCLUSION:The FCIA for detecting plasma vWF:Ag is not only an effective supplement to ELISA, but also the efficiency is faster and more sensitive, thus improves the diagnosis of type 3 vWD. Elevated levels of vWF: Ag in IS patients indicate the poor recovery of daily activities and prognosis.
