title: Improved immunoassay for the determination of surfactant protein A (SP-A) in human amniotic fluid.
abstract: A simple, improved immunoassay for the determination of human surfactant protein A (SP-A) in human amniotic fluid was developed. The immunoreaction with a monoclonal antibody PC6-immobilized plastic bead, peroxidase-labeled monoclonal antibody PE10 and amniotic fluid sample diluting in the buffer containing 0.6% sodium dodecyl sulfate/2% Triton X-100, was carried out simultaneously at 45 degrees C for 30 min in test tubes. After washing the bead with 2% skimmilk in phosphate buffered saline containing 1% Triton X-100, the peroxidase reaction was developed by adding the substrate reagent and the absorbance was measured. The amniotic fluid obtained at full term was used as a standard instead of purified SP-A, because of the stability of the antigenicity. This immunoassay method was used to measure SP-A in 69 samples of amniotic fluid from 22 to 41 weeks of gestation. The result indicated that the SP-A values obtained by the present immunoassay can be used for predicting the fetal lung maturity. This simplified monoclonal immunoassay completed the measurement within 1 hr, and so it could be used routinely in clinical laboratory.
authors.affiliation: Department of Biochemistry, Sapporo Medical College.
authors.forename: H
authors.initials: H
authors.lastname: Shimizu
authors.forename: K
authors.initials: K
authors.lastname: Hosoda
authors.forename: M
authors.initials: M
authors.lastname: Mizumoto
authors.forename: Y
authors.initials: Y
authors.lastname: Kuroki
authors.forename: H
authors.initials: H
authors.lastname: Sato
authors.forename: K
authors.initials: K
authors.lastname: Kataoka
authors.forename: M
authors.initials: M
authors.lastname: Hagisawa
authors.forename: S
authors.initials: S
authors.lastname: Fujimoto
authors.forename: T
authors.initials: T
authors.lastname: Akino
mesh_terms: D000653:Amniotic Fluid / Q000032:analysis*; D000818:Animals; D001059:Apoproteins / Q000032:analysis*; D005260:Female; D005865:Gestational Age; D006801:Humans; D007118:Immunoassay; D007124:Immunoenzyme Techniques; D007231:Infant, Newborn; D051379:Mice; D008807:Mice, Inbred BALB C; D011247:Pregnancy; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000032:analysis*; D012127:Respiratory Distress Syndrome, Newborn / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate; D013696:Temperature
chemical_list: D001059:Apoproteins; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; C046616:pulmonary surfactant apoprotein; D012967:Sodium Dodecyl Sulfate