title: Negative interference by rheumatoid factor in alpha-fetoprotein chemiluminescent microparticle immunoassay.
abstract: Background Rheumatoid factor causes positive interference in multiple immunoassays. Recently, negative interference has also been found in immunoassays in the presence of rheumatoid factor. The chemiluminescent microparticle immunoassay is widely used to determine serum alpha-fetoprotein. However, it is not clear whether the presence of rheumatoid factor in the serum causes interference in the chemiluminescent microparticle immunoassay of alpha-fetoprotein. Methods Serum alpha-fetoprotein was determined using the ARCHITECT alpha-fetoprotein assay. The estimation of alpha-fetoprotein recovery was carried out in samples prepared by diluting high-concentration alpha-fetoprotein serum with rheumatoid factor-positive or rheumatoid factor-negative serum. Paramagnetic microparticles coated with hepatitis B surface antigen-anti-HBs complexes were used to remove rheumatoid factor from the serum. Results The average recovery of alpha-fetoprotein was 88.4% and 93.8% in the rheumatoid factor-positive and rheumatoid factor-negative serum samples, respectively. The recovery of alpha-fetoprotein was significantly lower in the rheumatoid factor-positive serum samples than in the rheumatoid factor-negative serum samples. In two of five rheumatoid factor-positive samples, a large difference was found (9.8%) between the average alpha-fetoprotein recoveries in the serially diluted and initial recoveries. Fourteen rheumatoid factor-positive serum samples were pretreated with hepatitis B surface antigen-anti-HBs complex-coated paramagnetic microparticles. The alpha-fetoprotein concentrations measured in the pretreated samples increased significantly. Conclusions It was concluded that the alpha-fetoprotein chemiluminescent microparticle immunoassay is susceptible to interference by rheumatoid factor, leading to significantly lower results. Eliminating the incidence of negative interference from rheumatoid factor should be an important goal for immunoassay providers. In the meantime, laboratorians must remain alert to the negative interference by rheumatoid factor, and in some cases, pretreat rheumatoid factor-positive samples with blocking or absorbing reagents.
authors.affiliation: 1 Department of Laboratory Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: 1 Department of Laboratory Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Bi
authors.affiliation: 1 Department of Laboratory Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: 2 Department of Laboratory Medicine, Zhongnan Hospital, Wuhan University, Wuhan, China.
authors.forename: Yirong
authors.initials: Y
authors.lastname: Li
mesh_terms: D016477:Artifacts; D006514:Hepatitis B Surface Antigens / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000592:standards*; D008163:Luminescent Measurements; D058185:Magnetite Nanoparticles; D011485:Protein Binding; D015203:Reproducibility of Results; D012217:Rheumatoid Factor / Q000097:blood*; D012680:Sensitivity and Specificity; D000509:alpha-Fetoproteins / Q000032:analysis*
chemical_list: D006514:Hepatitis B Surface Antigens; D058185:Magnetite Nanoparticles; D000509:alpha-Fetoproteins; D012217:Rheumatoid Factor
keywords: Rheumatoid factor; alpha-fetoprotein; immunoassay; negative interference