PMID- 14675557
OWN - NLM
STAT- MEDLINE
DCOM- 20040806
LR  - 20191210
IS  - 0009-9120 (Print)
IS  - 0009-9120 (Linking)
VI  - 37
IP  - 1
DP  - 2004 Jan
TI  - Development of an ultrasensitive enzyme immunoassay for human proadrenomedullin 
      N-terminal 20 peptide and direct measurement of two molecular forms of PAMP in 
      plasma from healthy subjects and patients with cardiovascular disease.
PG  - 14-21
AB  - OBJECTIVE: Proadrenomedullin N-terminal 20 peptide (PAMP) processed from an 
      adrenomedullin precursor is a potent hypotensive peptide. It was anticipated that 
      a mature form of PAMP (m-PAMP) and an intermediate PAMP-gly existed together in 
      the blood. To measure concentrations of PAMPs in human plasma directly, we have 
      developed a highly sensitive enzyme immunoassay (immune complex transfer enzyme 
      immunoassay, ICT-EIA). DESIGN AND METHODS: PAMP was reacted simultaneously with 
      2,4-dinitrophenyl (DNP)-biotinyl-bovine serum albumin (BSA)-anti-PAMP Fab' 
      conjugate and anti-PAMP Fab'-beta-D-galactosidase conjugate. The immune complex 
      that was formed was initially trapped onto a polystyrene bead coated with 
      anti-DNP IgG, and then transferred onto a second polystyrene bead coated with 
      streptavidin. The resulting three-component complex was then assayed 
      fluorometrically. RESULTS: The detection limits of ICT-EIA for both m-PAMP and 
      PAMP-gly were 0.1 pmol/l with as little as 10 microl of plasma, and were a 
      hundred times higher than with conventional radioimmunoassay (RIA). Using 
      ICT-EIA, we determined that the plasma concentrations of m-PAMP and PAMP-gly in 
      51 healthy volunteers were 0.51 +/- 0.19 and 1.15 +/- 0.38 pmol/l (mean +/- SD), 
      respectively. Both plasma m-PAMP and PAMP-gly concentrations in patients with a 
      variety of diseases, including hypertension, heart failure, chronic renal 
      failure, and hemodialysis, were significantly higher than those in healthy 
      subjects. In addition, both plasma m-PAMP and PAMP-gly concentrations in patients 
      with New York Heart Association (NYHA) class I-IV heart failure were increased in 
      proportion to clinical severity. CONCLUSIONS: These sensitive and specific 
      ICT-EIAs may be used as a powerful tool for investigating the cardiovascular 
      system in patients with heart failure.
FAU - Hashida, Seiichi
AU  - Hashida S
AD  - Department of Biochemistry, Miyazaki Medical College, Kiyotake, Miyazaki 
      889-1692, Japan. shashida@fc.miyazaki-med.ac.jp
FAU - Kitamura, Kazuo
AU  - Kitamura K
FAU - Nagatomo, Yoshitatsu
AU  - Nagatomo Y
FAU - Shibata, Yoshisato
AU  - Shibata Y
FAU - Imamura, Takuroh
AU  - Imamura T
FAU - Yamada, Kazuhiro
AU  - Yamada K
FAU - Fujimoto, Shouichi
AU  - Fujimoto S
FAU - Kato, Johji
AU  - Kato J
FAU - Morishita, Kazuhiro
AU  - Morishita K
FAU - Eto, Tanenao
AU  - Eto T
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Clin Biochem
JT  - Clinical biochemistry
JID - 0133660
RN  - 0 (Immunoglobulin Fab Fragments)
RN  - 0 (Peptides)
RN  - 148498-78-6 (Adrenomedullin)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - Adrenomedullin
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Animals
MH  - Calibration
MH  - Cardiovascular Diseases/*blood
MH  - Cross Reactions
MH  - Female
MH  - Humans
MH  - *Immunoenzyme Techniques
MH  - Immunoglobulin Fab Fragments/immunology
MH  - Male
MH  - Middle Aged
MH  - Peptides/*blood/immunology
MH  - Rats
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - beta-Galactosidase/analysis
EDAT- 2003/12/17 05:00
MHDA- 2004/08/07 05:00
CRDT- 2003/12/17 05:00
PHST- 2003/12/17 05:00 [pubmed]
PHST- 2004/08/07 05:00 [medline]
PHST- 2003/12/17 05:00 [entrez]
AID - S0009912003001607 [pii]
AID - 10.1016/j.clinbiochem.2003.09.007 [doi]
PST - ppublish
SO  - Clin Biochem. 2004 Jan;37(1):14-21. doi: 10.1016/j.clinbiochem.2003.09.007.