return (home page)
return (main kit list) return (lipoprotein
antibodies index)
rev: November 26, 2007
MICROFLUORALä
Microalbumin Test
A Quick, Quantitative,
Fluorescence Test for the Detection of Microalbuminuria
Cat. No.: RDI-PROPR2005 $495.00/kit
Size of the Kit: 5 x 96
Determinations
Storage: 2 - 8° C
1.
Clinical Significance
Persistent
microalbuminuria, defined as a urinary excretion rate
of 20 - 200 micrograms/ml, indicates a high probability of damage of the glomerular filtration capacity of the kidney and is of
great diagnostic relevance
·
in diabetic patients for early diagnosis of nephropathy
·
in hypertensive patients as indicator of end-organ damage associated
with lower life expectancy,
·
in pregnancy as predictor of delivering preeclampsia,
and
·
is probably also associated with cardiovascular disease
in non-diabetic subjects.
All
patients with potential disease involvement of the kidneys should be screened
for early diagnosis.
2. Application
Microfluoral test is a quick
low-cost method with high accuracy for rapid quantification of microalbuminuria
in large sample numbers.
3. Test Principle
The assay is based
on Albumin Blue 580 (AB580) and performed in a 96-well microplate
and measured in a Fluorescence Microplate Reader at
590-600 nm excitation and 630-645 nm emission. AB580 binds to albumin by
forming a strongly fluorescent complex without interfering with urinary
proteins or frequently used drugs. Sample and reagent are pipetted
into the wells of a microplate. The plate can be
measured without further incubation. Fluorescence intensity directly reflects
albumin concentration. A set of standards gives an almost linear standard curve
from 0 to 200 mg/l. Detection limit is 2mg/l albumin.
4. Material Required
· Precision pipettes
· Sterile pipette tips
Microplate mixer
or 8-channel multipipette
· Fluorescence microplate reader with
excitation filter 590-600 nm and emission filter 630-645 nm
5. Contents of
Test Kit
12x8-well microplates,
5 pcs.
P Positive
control, 500 µl (ready-to-use). Contains human albumin*.
S1 Human albumin* standard 200
mg/l, 500 µl (ready-to-use).
S2 Human albumin* standard 100 mg/l,
500 µl (ready-to-use).
S3 Human albumin* standard 30
mg/l, 500 µl (ready-to-use).
S4 Human albumin* standard 2
mg/l, 500 µl (ready-to-use).
Dil Dilution
buffer**, 2x 50 ml
Dye AB580 concentrate*** (50x), 2.5 ml
* Human albumin was found negative for HIV I und II, Hepatitis B and C.
Nevertheless, standards and control should be handled as potentially infectious
material.
** Standards, control and buffer contain sodium azide
as preservative.
*** Contains Isopropanol, flammable and
irritant.
6. Specimen and
Sample Storage
Random urine is
the simplest approach to obtaining a specimen for analysis, but gives the most
variable results. The preferred specimen is 24-hr urine without preservative.
The albumin measurement is multiplied by the volume expressed in L to obtain mg
albumin excreted in 24 hr. Refrigerated samples are stable for up to two weeks.
Samples may be stored at -20°C for up to 6 months.
7. Preparation
Warm
up kit to room temperature. Just before use prepare the amount of AB580 working
reagent necessary and mix well:
For 1x 8-well microplate
strip: 30µl AB580- concentrate ad
1500µl dilution buffer
For 1x 96well microplate:
360 µl AB580 concentrate ad 18ml dilution
buffer
Attention: Use polypropylene vials
only!
7. Test Procedure
1.
Place 25 µl standard S1 to
S4, positive control and specimen in appropriate wells of the microplate.
2.
Add 150µl AB580 working
reagent.
3.
Mix well, e.g. by suspending
with an 8-channel multipipette or by mixing in a microplate mixer.
4.
Measure fluorescence at
590-600 nm excitation and 630-645 nm emission in a fluorescence microplate Reader.
9. Calculation
and Interpretation of Results
Read Relative Fluorescence Counts (RFC) of specimen
from the standard curve.
Example of a standard curve:
|
<20
mg/l
<30 mg/die
|
normal
no indication of microalbuminuria
|
|
20-200 mg/l
30-300 mg/die
|
pathological:
microalbuminuria
|
|
> 200 mg/l
> 300 mg/die
|
albuminuria
|