Lab Med Online.  2012 Apr;2(2):87-94.

Evaluation of a Modified Enzyme Linked Immunosorbent Assay for Serum PIVKA-II Measurement

• Park CH
• Seo JY
• Kim SY
• Jeagal TY
• Kang ES

Abstract

BACKGROUND
The serum des-gamma-carboxyprothrombin (protein induced by vitamin K antagonist-II, PIVKA-II) is a useful tumor marker in addition to alpha-fetoprotein for diagnosing primary hepatocellular carcinoma (HCC). In this study, we evaluated the laboratory performance of a modified ELISA method for PIVKA-II measurement adopting an automated ELISA processor in comparison with conventional manual method and investigated its diagnostic performance in patients with HCC.
METHODS
The laboratory performance of modified ELISA using PIVKA-II ELISA kit (Sanko Junyaku Co., Japan) was evaluated using control materials (10, 25, 500, 1,000 mAU/mL) and 208 patient samples according to the CLSI guidelines. In 93 HCC patients and 88 disease controls (30 chronic hepatitis and 58 liver cirrhosis), ROC curve, sensitivity, specificity, and positive and negative predictive values were analyzed.
RESULTS
Total and within-run CVs for middle, high and very high level samples were less than 10%, while those of low level samples were over 10% (12.6% and 11.7%, respectively). The modified ELISA showed an excellent linearity (r>0.99) and low carryover rate (-0.14%). Although the correlation between the conventional and modified ELISAs was excellent (r=0.982), there was a proportional deviation of PIVKA-II levels (y intercept: 0.621). With a cut-off of 30 mAU/mL, the sensitivity and specificity of PIVKA-II for the diagnosis of HCC were 58% and 92%, respectively.
CONCLUSIONS
PIVKA-II measurement by modified ELISA using an automated ELISA processor can improve the efficiency of laboratory in terms of turnaround time and labor intensiveness while maintaining reasonable sensitivity and specificity for the diagnosis of HCC.

Keyword

PIVKA-II; Alpha-fetoprotein; Immunoassay; Hepatocellular carcinoma

MeSH Terms

alpha-Fetoproteins
Biomarkers
Carcinoma, Hepatocellular
Enzyme-Linked Immunosorbent Assay
Hepatitis, Chronic
Humans
Immunoassay
Liver
Protein Precursors
Prothrombin
ROC Curve
Sensitivity and Specificity
Vitamin K
Protein Precursors
Prothrombin
Vitamin K
alpha-Fetoproteins

Figure

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  Fig. 1 Linearity of PIVKA-II measurement by modified ELISA method.

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  Fig. 2 Comparison of the PIVKA-II value between conventional and modified ELISA methods.

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  Fig. 3 Bland-Altman plots of low group (A), middle group (B), high group (C), very high group (D) of serum PIVKA-II between conventional and modified ELISA methods.

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  Fig. 4 Box plots comparing serum levels of biomarkers according to the size of HCC. For AFP, the log data are presented in order to accommodate the wide range. The box refers to the 25th and 75th percentile values with a line indicating the median levels, whereas the interquartile range extends outside the box. The mean of three groups, according to different size of tumor, is significantly different for PIVKA-II (P=0.020, Kruskal-Wallis test) (A) but not for AFP (P=0.106, Kruskal-Wallis test) (B). Statistical significance between two different groups are indicated with symbols (# if P<0.05, ## if P≥0.05, Independent T test or Mann-Whitney U test). Data (box plots); Mild outliers (circles); Extreme outliers (asterisk).

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  Fig. 5 ROC curve evaluating cases with hepatocellular carcinoma and cirrhosis. The area under the curve is shown with its 95% confidence intervals.

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