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Annals of Clinical & Laboratory Science 31:205-210 (2001)
© 2001 Association of Clinical Scientists

Use of Microtiter Plates for Latex Agglutination Immunoassay of Serum Alpha2-Macroglobulin

Tetsuo Makino
Department of Clinical Chemistry, Kanagawa Prefectural College of Nursing and Medical Technology, Yokohama, Japan

Address correspondence to Tetsuo Makino, Ph.D., Department of Clinical Chemistry, Kanagawa Prefectural College of Nursing and Medical Technology, 1-5-1 Nakao, Asahi-ku, Yokohama 241-0815, Japan; tel 81 045 361 6141; fax 81 045 362 8785; e-mail dr_tetsu{at}xb3.so-net.ne.jp

A simple, sensitive turbidimetric immunoassay is described for analysis of alpha2-macroglobulin (AMG) in serum, using microtiter plates and a commercial kit that features a sensitive latex reagent. Using microwells as reaction vessels and the latex reagent made it possible to minimize the sample size and the amounts of reagents. The procedure involves two pipetting steps and a brief incubation (10 min) at room temperature; the assay measures AMG concentrations ranging from 0.12 to 5 g/L and is unaffected by icterus, hemolysis, lipemia, or rheumatoid factor, even at high concentrations. Recovery of AMG added to three pooled serum samples averaged 95–105%. CVs for replicate analyses ranged from 1.9 to 4.8% (within-run) and 6.1 to 9.3% (between-run). Based on paired analyses of 70 serum specimens, AMG concentrations obtained by this turbidimetric microtiter immunoassay were in close agreement (correlation coefficient = 0.99) with results obtained by an enzyme-linked immunosorbent assay (ELISA). Serum AMG concentrations in 48 healthy adults (mean ± 2SD) were 1.87 ± 1.00 g/L.

Keywords: Alpha2-macroglobulin, latex immunoassay, turbidimetry, microtiter plates

Abbreviations: AMG, alpha2-macroglobulin; SRID, single radial immunodiffusion; RIE, rocket immunoelectrophoresis; ELISA, enzyme-linked immunosorbent assay; TIA, turbidimetric immunoassay; PEG, polyethylene glycol; BSA, bovine serum albumin






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