Establishment of a New Protein C Detection System Based on Chromogenic Substrate Assay and Its Clinical Diagnostic Value for Deep Vein Thrombosis

Overview

Journal J Clin Lab Anal

Publisher Wiley

Specialties Biomedical Engineering
Biotechnology
Pathology

Date 2021 Nov 13

PMID 34773713

Citations 1

Authors

Wenfei Lu

Anshan Ge

Jinxia Zhao

Hui Rong

Chao Zhu

Limin Lun

Affiliations

Soon will be listed here.

Abstract

Background: Deficiency of protein C (PC) affects the balance between blood coagulation and fibrinolysis in the human body. Chromogenic-based assay is recommended as the preferred screening method for detecting PC deficiency. We established a PC detection system based on the chromogenic substrate assay.

Methods: First, a kit for the determination of PC activity in plasma was elaborately developed and its reaction parameters on XL-3200c were explored. Then, we evaluated its performance and collected specimens to compare the test results obtained with those of the Siemens detection system. Finally, the clinical diagnostic efficacy of this detection system for deep vein thrombosis (DVT) was assessed.

Results: Optimum conditions for PC detection were 0.25-0.1 U/ml protein C activator Protac and 2.5-1 mM Pefachrome PCa5297. The composition and concentration ranges of buffer substances and stabilizers in the kit were also explored. Satisfactory results were observed in performance evaluation. The test results of the newly built detection system were highly correlated with those of the Siemens detection system (R = 0.9771 in the control group and R = 0.9776 in the DVT group), and Bland-Altman plots also showed high consistency between the two detection systems. In addition, the area under the curve (AUC) of the newly built PC detection system for DVT was 0.888, indicating this system could effectively improve the diagnostic sensitivity and specificity for DVT.

Conclusion: In this study, a sensitive, wide linear range and reliable PC activity detection system were established.

Citing Articles

Establishment of a new protein C detection system based on chromogenic substrate assay and its clinical diagnostic value for deep vein thrombosis.

Lu W, Ge A, Zhao J, Rong H, Zhu C, Lun L J Clin Lab Anal. 2021; 35(12):e24109.

PMID: 34773713 PMC: 8649361. DOI: 10.1002/jcla.24109.

References

Raizman J, Tsui A, Goudreau B, Fuzery A, Estey M, Sadrzadeh H . Multi-platform analytical evaluation of the Beckman Coulter Access high-sensitivity troponin I assay across different laboratory sites using Barricor plasma. Clin Biochem. 2019; 78:25-31. DOI: 10.1016/j.clinbiochem.2019.10.014. View

Weiler H . Regulation of inflammation by the protein C system. Crit Care Med. 2010; 38(2 Suppl):S18-25. DOI: 10.1097/CCM.0b013e3181c9cbb5. View

Martinez-Morillo E, Diamandis A, Diamandis E . Reference intervals and biological variation for kallikrein 6: influence of age and renal failure. Clin Chem Lab Med. 2012; 50(5):931-4. DOI: 10.1515/cclm-2012-0075. View

Baglin T, Gray E, Greaves M, Hunt B, Keeling D, Machin S . Clinical guidelines for testing for heritable thrombophilia. Br J Haematol. 2010; 149(2):209-20. DOI: 10.1111/j.1365-2141.2009.08022.x. View

Martinez-Iribarren A, Tejedor X, Sala Sanjaume A, Leis A, Dolade Botias M, Morales-Indiano C . Performance evaluation of the new hematology analyzer UniCel DxH 900. Int J Lab Hematol. 2021; 43(4):623-631. DOI: 10.1111/ijlh.13448. View

Adcock Funk D . Coagulation assays and anticoagulant monitoring. Hematology Am Soc Hematol Educ Program. 2012; 2012:460-5. DOI: 10.1182/asheducation-2012.1.460. View

Volz K, Buhler B, Muller C . Protein C activators from snake venoms and their diagnostic use. Haemostasis. 2002; 31(3-6):266-72. DOI: 10.1159/000048072. View

Marlar R, Gausman J . Laboratory testing issues for protein C, protein S, and antithrombin. Int J Lab Hematol. 2014; 36(3):289-95. DOI: 10.1111/ijlh.12219. View

Lu W, Ge A, Zhao J, Rong H, Zhu C, Lun L . Establishment of a new protein C detection system based on chromogenic substrate assay and its clinical diagnostic value for deep vein thrombosis. J Clin Lab Anal. 2021; 35(12):e24109. PMC: 8649361. DOI: 10.1002/jcla.24109. View

10.

Liu H, Wang H, Tang L, Yang Y, Wang Q, Zeng W . Compound heterozygous protein C deficiency in a family with venous thrombosis: Identification and in vitro study of p.Asp297His and p.Val420Leu mutations. Gene. 2015; 563(1):35-40. DOI: 10.1016/j.gene.2015.03.002. View

11.

Angchaisuksiri P, Atichartakarn V, Aryurachai K, Archararit N, Rachakom B, Atamasirikul K . Risk factors of venous thromboembolism in thai patients. Int J Hematol. 2008; 86(5):397-402. DOI: 10.1007/BF02983995. View

12.

Gu Y, Shen W, Zhang L, Zhang J, Ying C . Deficiency of antithrombin and protein C gene in 202 Chinese venous thromboembolism patients. Int J Lab Hematol. 2013; 36(2):151-5. DOI: 10.1111/ijlh.12146. View

13.

Goldenberg N, Manco-Johnson M . Protein C deficiency. Haemophilia. 2009; 14(6):1214-21. DOI: 10.1111/j.1365-2516.2008.01838.x. View

14.

Stocker K, Fischer H, Meier J, Brogli M, Svendsen L . Characterization of the protein C activator Protac from the venom of the southern copperhead (Agkistrodon contortrix) snake. Toxicon. 1987; 25(3):239-52. DOI: 10.1016/0041-0101(87)90253-4. View

15.

Fushimi Y, Tatebe J, Okuda Y, Ishii T, Ujiie S, Morita T . Performance evaluation of an Indoxyl Sulfate Assay Kit "NIPRO". Clin Chem Lab Med. 2019; 57(11):1770-1776. DOI: 10.1515/cclm-2019-0218. View

16.

Khor B, Van Cott E . Laboratory tests for protein C deficiency. Am J Hematol. 2010; 85(6):440-2. DOI: 10.1002/ajh.21679. View

17.

Lin Z, Fang Y, Jin H, Lin H, Dai Z, Luo Q . Performance evaluation of a chemiluminescence microparticle immunoassay for CK-MB. J Clin Lab Anal. 2018; 32(6):e22426. PMC: 6816894. DOI: 10.1002/jcla.22426. View

18.

Li H, Wong B, Ip W, Yeung W, Ho P, Ng E . Comparative evaluation of three new commercial immunoassays for anti-Müllerian hormone measurement. Hum Reprod. 2016; 31(12):2796-2802. DOI: 10.1093/humrep/dew248. View

19.

Connors J . Thrombophilia Testing and Venous Thrombosis. N Engl J Med. 2017; 377(12):1177-1187. DOI: 10.1056/NEJMra1700365. View

20.

Tamura S, Suga Y, Tanamura M, Murata-Kawakami M, Takagi Y, Hottori Y . Optimisation of antithrombin resistance assay as a practical clinical laboratory test: Development of prothrombin activator using factors Xa/Va and automation of assay. Int J Lab Hematol. 2018; 40(3):312-319. DOI: 10.1111/ijlh.12786. View