Optimisation of antithrombin resistance assay as a practical clinical laboratory test: Development of prothrombin activator using factors Xa/Va and automation of assay

Tamura, Shogo; Suga, Yanto; Tanamura, M.; Murata-Kawakami, Moe; Takagi, Yuki; Hottori, Y.; Kakihara, Misaki; Suzuki, Sachiko; Takagi, Akira; Kojima, Tetsuhito

doi:10.1111/ijlh.12786

Cited by 3 publications

(3 citation statements)

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“…The correlation equation and correlation coefficient were determined, and the consistency of the two detection systems was presented with the Bland‐Altman plot. In addition, receiver operating characteristic (ROC) curves were used to analyze and compare the diagnostic efficiency of the two detection systems for DVT 13,14 …”

Section: Methodsmentioning

confidence: 99%

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

Ge²,

Zhao

et al. 2021

Clinical Laboratory Analysis

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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 (R2 = 0.9771 in the control group and R2 = 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.

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Section: Methodsmentioning

confidence: 99%

“…In addition, receiver operating characteristic (ROC) curves were used to analyze and compare the diagnostic efficiency of the two detection systems for DVT. 13 , 14 …”

Section: Methodsmentioning

confidence: 99%

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

Ge²,

Zhao

et al. 2021

Clinical Laboratory Analysis

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“…Phenotypic assays have been described for AT resistance, but they are not commercially available. 224 227…”

Section: Other Thrombophiliasmentioning

confidence: 99%

Thrombophilia Screening: Not So Straightforward

Moore

2024

Semin Thromb Hemost

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Although inherited thrombophilias are lifelong risk factors for a first thrombotic episode, progression to thrombosis is multifactorial and not all individuals with inherited thrombophilia develop thrombosis in their lifetimes. Consequently, indiscriminate screening in patients with idiopathic thrombosis is not recommended, since presence of a thrombophilia does not necessarily predict recurrence or influence management, and testing should be selective. It follows that a decision to undertake laboratory detection of thrombophilia should be aligned with a concerted effort to identify any significant abnormalities, because it will inform patient management. Deficiencies of antithrombin and protein C are rare and usually determined using phenotypic assays assessing biological activities, whereas protein S deficiency (also rare) is commonly detected with antigenic assays for the free form of protein S since available activity assays are considered to lack specificity. In each case, no single phenotypic assay is capable of detecting every deficiency, because the various mutations express different molecular characteristics, rendering thrombophilia screening repertoires employing one assay per potential deficiency, of limited effectiveness. Activated protein C resistance (APCR) is more common than discrete deficiencies of antithrombin, protein C, and protein S and also often detected initially with phenotypic assays; however, some centres perform only genetic analysis for factor V Leiden, as this is responsible for most cases of hereditary APCR, accepting that acquired APCR and rare F5 mutations conferring APCR will go undetected if only factor V Leiden is evaluated. All phenotypic assays have interferences and limitations, which must be factored into decisions about if, and when, to test, and be given consideration in the laboratory during assay performance and interpretation. This review looks in detail at performance and limitations of routine phenotypic thrombophilia assays.

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Blood coagulation regulation and thrombosis

Kojima¹

2020

Nihon Kessen Shiketsu Gakkaishi

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Optimisation of antithrombin resistance assay as a practical clinical laboratory test: Development of prothrombin activator using factors Xa/Va and automation of assay

Abstract: We optimised the ATR assay with the FXa/Va activator and adapted the assay for ACL TOP 500; the assay showed the ability to clearly detect antithrombin-resistant prothrombin in manual and automated procedures.

Cited by 3 publications

References 18 publications

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

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

Thrombophilia Screening: Not So Straightforward

Blood coagulation regulation and thrombosis

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