A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids.
We describe here the development of a chemifluorescent competitive enzyme-linked immunosorbent assay (ELISA) that quantifies marinobufagenin (MBG) levels in biological fluids. Based on a polyclonal antibody raised against a novel MBG-bovine serum albumin conjugate, this assay achieved an MBG detection limit of less than 9 pg/mL. MBG levels in various rat urine and serum samples were effectively determined using this methodology. Interassay variability averaged 9.8%, while intra-assay variability averaged 1.9 and 2.5% in representative serum and urine samples, respectively. Recovery of exogenously added MBG averaged 106%, and parallelism data further established the accuracy of the assay. Employment of this assay to detect MBG abnormalities represents a powerful tool for the possible diagnosis, prevention and management of human hypertensive states, particularly preeclampsia.
Department of Poultry Science, Texas A&M University, College Station, Texas, USA.
Daad
D
Abi-Ghanem
Xinzhong
X
Lai
Luc R
LR
Berghman
Darijana
D
Horvat
Jing
J
Li
Daniel
D
Romo
Mohammad N
MN
Uddin
Yoshiaki
Y
Kamano
Toshihiko
T
Nogawa
Jun-Ping
JP
Xu
George R
GR
Pettit
Jules B
JB
Puschett
D000818:Animals; D002018:Bufanolides / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D011225:Pre-Eclampsia / Q000175:diagnosis; D011247:Pregnancy; D011817:Rabbits; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry
D002018:Bufanolides; D012710:Serum Albumin, Bovine; C093896:marinobufagenin