Development and application of a fluorescence protein microarray for detecting serum alpha-fetoprotein in patients with hepatocellular carcinoma

J Int Med Res. 2016 Dec;44(6):1414-1423. doi: 10.1177/0300060516672370. Epub 2016 Nov 24.

Abstract

Objective To develop a simple, effective, time-saving and low-cost fluorescence protein microarray method for detecting serum alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC). Method Non-contact piezoelectric print techniques were applied to fluorescence protein microarray to reduce the cost of prey antibody. Serum samples from patients with HCC and healthy control subjects were collected and evaluated for the presence of AFP using a novel fluorescence protein microarray. To validate the fluorescence protein microarray, serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA). Results A total of 110 serum samples from patients with HCC ( n = 65) and healthy control subjects ( n = 45) were analysed. When the AFP cut-off value was set at 20 ng/ml, the fluorescence protein microarray had a sensitivity of 91.67% and a specificity of 93.24% for detecting serum AFP. Serum AFP quantified via fluorescence protein microarray had a similar diagnostic performance compared with ELISA in distinguishing patients with HCC from healthy control subjects (area under receiver operating characteristic curve: 0.906 for fluorescence protein microarray; 0.880 for ELISA). Conclusion A fluorescence protein microarray method was developed for detecting serum AFP in patients with HCC.

Keywords: Alpha-fetoprotein; detecting; fluorescence protein microarray; hepatocellular carcinoma.

MeSH terms

  • Aged
  • Antibodies / chemistry
  • Area Under Curve
  • Biomarkers, Tumor / blood
  • Biomarkers, Tumor / genetics*
  • Carbocyanines / chemistry
  • Carcinoma, Hepatocellular / blood
  • Carcinoma, Hepatocellular / diagnosis*
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / pathology
  • Case-Control Studies
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Fluorescence
  • Fluorescent Dyes / chemistry
  • Gene Expression
  • Humans
  • Liver Neoplasms / blood
  • Liver Neoplasms / diagnosis*
  • Liver Neoplasms / genetics
  • Liver Neoplasms / pathology
  • Male
  • Middle Aged
  • Protein Array Analysis / methods*
  • Protein Array Analysis / standards
  • ROC Curve
  • Streptavidin / chemistry
  • alpha-Fetoproteins / genetics*
  • alpha-Fetoproteins / metabolism

Substances

  • Antibodies
  • Biomarkers, Tumor
  • Carbocyanines
  • Fluorescent Dyes
  • alpha-Fetoproteins
  • cyanine dye 3
  • Streptavidin