Direct Immunochemiluminescent Assay for proBNP and Total BNP in Human Plasma proBNP and Total BNP Levels in Normal and Heart Failure.

• Nishikimi, Toshio

• Okamoto, Hiroyuki

• Nakamura, Masahiro

• Ogawa, Naoko

• Horii, Kazukiyo

• Nagata, Kiyoshi

• Nakagawa, Yasuaki

• Kinoshita, Hideyuki

• Yamada, Chinatsu

• Nakao, Kazuhiro

• Minami, Takeya

• Kuwabara, Yoshihiro

• Kuwahara, Koichiro

• Masuda, Izuru

• Kangawa, Kenji

• Minamino, Naoto

• Nakao, Kazuwa

Abstract

[Background]Recent studies have shown that in addition to brain (or B-type) natriuretic peptide (BNP) and the N-terminal proBNP fragment, levels of intact proBNP are also increased in heart failure. Moreover, present BNP immunoassays also measure proBNP, as the anti-BNP antibody cross-reacts with proBNP. It is important to know the exact levels of proBNP in heart failure, because elevation of the low-activity proBNP may be associated with the development of heart failure. [Methodology/Principal Findings]We therefore established a two-step immunochemiluminescent assay for total BNP (BNP+proBNP) and proBNP using monoclonal antibodies and glycosylated proBNP as a standard. The assay enables measurement of plasma total BNP and proBNP within only 7 h, without prior extraction of the plasma. The detection limit was 0.4 pmol/L for a 50-µl plasma sample. Within-run CVs ranged from 5.2%–8.0% in proBNP assay and from 7.0%–8.4% in total BNP assay, and between-run CVs ranged from 5.3–7.4% in proBNP assay and from 2.9%–9.5% in total BNP assay, respectively. The dilution curves for plasma samples showed good linearity (correlation coefficients = 0.998–1.00), and analytical recovery was 90–101%. The mean total BNP and proBNP in plasma from 116 healthy subjects were 1.4±1.2 pM and 1.0±0.7 pM, respectively, and were 80±129 pM and 42±70 pM in 32 heart failure patients. Plasma proBNP levels significantly correlate with age in normal subjects. [Conclusions/Significance]Our immunochemiluminescent assay is sufficiently rapid and precise for routine determination of total BNP and proBNP in human plasma.

Journal

• PloS one

  PloS one 8 (1), 2013-01-24

  Public Library of Science

Details

• CRID: 1050564285715186560
• NII Article ID: 120005241437
• ISSN: 19326203
• HANDLE: 2433/169792
• Text Lang: en
• Article Type: journal article
• Data Source:
  • IRDB
  • CiNii Articles