Abstracts of the Scientific Posters, 2013 AACC Annual Meeting ...

Technology/Design Development
Wednesday, July 31, 9:30 am – 5:00 pm
Samples: The serum and plasma samples were collected from inpatients/outpatients
and the employees who volunteered. This study has been approved by the ethical
committee in Hamamatsu University School of Medicine.
Material and Method: The reagent PANACLEAR MMP-3 “Latex” (Sekisui Medical
Co., Ltd.) and the automated clinical chemistry analyzer LABOSPECT 008 (Hitachi)
were used for the following evaluations: (1) precision, (2) dilution linearity (2 level of
MMP-3 samples were diluted with physiological saline) and limit of detection (2.6SD
method), (3) interference (evaluated with Interference Check (Sysmex) and ascorbic
acid), (4) correlation with results from JCA-BM1650 (JEOL Ltd.), and (5) probe
contamination test (evaluated on 42 parameters of the same pre-installed module).
Results: (1) Within-run precision of CV (n=20) :1.35 % (Control L; mean 110.9 ng/
mL), 0.78 % (Control H; mean 435.4 ng/mL ), 1.62 % (pooled serum; mean 99.2 ng/
mL) (2) Linearity was up to 1500 ng/mL and limit of detection was 9.85 ng/mL. (3)
No influences were observed by bilirubin < 200 mg/L, hemoglobin < 5 g/L, RF < 550
U/mL or ascorbic acid < 500 mg/L in sample. (4) High correlation was noted, with the
regression formula being y = 0.991x-13.9 and the correlation coefficient being 0.985
(N=92). (5) Probe contamination test: No influence by prove contamination test was
noted on any of the 42 parameters.
Conclusion: PANACLEAR MMP-3 “Latex” with LABOSPECT 008 was shown in
the present study as having favorable performance. Wide assay range with linearity
up to 1500 ng/ml is especially convenient because high level MMP-3 samples don’t
require dilution and retesting. Additionally, in couple with LABOSPECT 008, routine
measurement of MMP-3 could be easily handled, and real-time reporting could lead
to clinical remission. This reagent could be useful for measuring MMP-3 in clinical
laboratories.
B-255
GlycA and GlycB: Novel NMR Markers of Systemic Inflammation
J. Otvos, I. Shalaurova, J. Wolak-Dinsmore, S. Matyus. LipoScience,
Raleigh, NC
Background: Serum concentrations of many glycosylated acute-phase proteins such
as C-reactive protein (CRP) and fibrinogen are used clinically to assess and monitor
both acute and chronic inflammation. Serum NMR spectra obtained for lipoprotein
particle analysis using the automated NMR Profiler system contain two NMR signals
originating from N-acetyl methyl group protons on the N- and O-linked glycans of
serum glycoproteins. One, that we named GlycA, comes from N-acetylglucosamine
and N-acetylgalactosamine and the other, GlycB, from sialic acid moieties. We
hypothesized that the measured amplitudes of these signals would reflect global
protein glycosylation levels, thereby providing measures of inflammation status
that might have clinical utility similar or complementary to existing inflammatory
biomarkers.
Methods: We used archived serum NMR spectra from previously-performed
automated NMR LipoProfile (lipoprotein particle) analyses conducted using the 400
MHz NMR Profiler analyzers at LipoScience. As shown in the Figure, the GlycA
and GlycB signals overlap the complex signal envelope from the allylic protons
of the lipids in VLDL, LDL, and HDL particles. To accurately quantify their
signal amplitudes, we developed an automated linear non-negative least-squares
deconvolution algorithm that takes account of the spectral contributions of serum
protein and 59 different lipoprotein subclasses. GlycA and GlycB levels are reported
in units of μmol/L N-acetyl methyl groups.
Results: GlycA and GlycB levels were measured with good precision (0.97). With the
automation the %CVs of all three concentration levels were