Korean J Lab Med.  2009 Oct;29(5):415-422. 10.3343/kjlm.2009.29.5.415.

Performance Evaluation of Affinity Column Mediated Immunometric Assay for Tacrolimus

• Chung JW
• An D
• Song J
• Chung HJ
• Park HI
• Lee W
• Chun S
• Min WK

• KMID: 1077417
• DOI: http://doi.org/10.3343/kjlm.2009.29.5.415

Abstract

BACKGROUND
Therapeutic drug monitoring (TDM) of tacrolimus is essential because of narrow therapeutic range and poor correlation of dose to blood concentration. Affinity Column Mediated Immunometric Assay (ACMIA) does not require a pretreatment steps in measurement of tacrolimus. In this study, we evaluated the performance of tacrolimus assay using ACMIA (Dimension RxL Max, Dade Behring).
METHODS
The imprecision, the linearity and the detection limits and the interferences by bilirubin and chyle, and correlation with hematocrit for tacrolimus by ACMIA were evaluated according to Clinical and Laboratory Standards Institute guidelines EP5-A2, EP6-A, EP17-A, EP9-A2, and EP7-A2. Method comparison studies with microparticle enzyme immunoassay (MEIA) (IMx Tacrolimus II, Abbott Laboratories) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) (Waters 2795 Quattromicro API, Micromass) were also performed.
RESULTS
The total imprecision for low, middle and high level was 12.8%, 9.0% and 6.7%, respectively. The range of tacrolimus from 3.1 ng/mL to 35.4 ng/mL showed a clinically relevant linearity. The limit of detection and the functional sensitivity were 0.24 ng/mL and 0.72 ng/mL, respectively. Tacrolimus concentration measurement (Tac-CM) with ACMIA did not show significant interferences with bile and chyle and also did not show significant correlation with hematocrit. In comparison study for Tac-CM with MEIA and LC-MS/MS, Tac-CM with ACMIA showed a good correlation with MEIA (r=0.950) and LC-MS/MS (r=0.946).
CONCLUSIONS
The imprecision, linearity, detection limits, interference and correlation of Tac-CM with ACMIA were suitable for clinical use. Tac-CM with ACMIA could reduce turn around time and help clinicians to manage transplant patients on immunosuppressant therapy.

Keyword

Tacrolimus; Affinity column mediated immunometric assay; Therapeutic drug monitoring

MeSH Terms

Bilirubin/chemistry
Chromatography, Affinity
Chyle/chemistry
Drug Monitoring
Humans
Immunoassay/*methods
Immunosuppressive Agents/*blood
Limit of Detection
Reagent Kits, Diagnostic
Reproducibility of Results
Tacrolimus/*blood

Figure

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  Fig. 1. Linearity of ACMIA for tacrolimus. Abbreviation: See Table 1.

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  Fig. 2. The functional sensitivity of tacrolimus measurement using ACMIA. Abbreviation: See Table 1.

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  Fig. 3. Comparisons of tacrolimus measurements among instruments (n=50, respectively). (A) IMx (MEIA) versus Dimension (ACMIA) using Deming regression analysis. (B) Bland-Altman analysis between ACMIA and MEIA, (C) LC-MS/MS versus ACMIA using Deming regression analysis, (D) Bland-Altman analysis between ACMIA and LC-MS/MS, (E) LC-MS/MS versus MEIA using Deming regression analysis and (F) Bland-Altman analysis between MEIA and LC-MS/MS. Abbreviations: MEIA, microparticle enzyme immunoassay; ACMIA, affinity column mediated immunometric assay; LC-MS/MS, liquid chromatography-tandem mass spectrometry.

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  Fig. 4. Relationship between hematocrit and tacrolimus concentrations. Group 1, target concentration 7.5 ng/mL (y=0.019x+6.624, r2=0.178, P=0.118); group 2, target concentration 20.0 ng/mL (y= 0.011x+18.675, r2=0.059, P=0.344). Abbreviation: Hct, hematocrit.

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