UMA CO., LTD.

MEASURE NAG
2-19-6 Yokosuka Reagent for measuring N-acetyl-β-D-glucosaminidase
Matsudo, Chiba, Japan MPT-NAG Substrate

 2 ~ 8 °C IVD In vitro Diagnostics Packages

R1 1  60 mL R2 1  20 mL
 DO NOT freeze  18 months/block from light R1 1  45 mL R2 1  15 mL

1. PURPOSE OF USE 7. STANDARD MEASUREMENT OPERATION

In vitro Determination of N-acetyl-β-D-glucosaminidase Specimen Calibrator Blank
(NAG) in urine. (S) (Std) (B)

2. GENERAL INSTRUCTION Specimen (L) 130 - -

1. For in vitro diagnostics use only. Calibrator (L) - 130 -

2. Diagnosis should be made in a comprehensive manner, Saline (L) - - 130

in accordance with other related test results and clinical R-1 (L) 1500 1500 1500

symptoms by the doctor in attendance. Incubate at 37 C in 5 minutes

3. For guaranteed results, usage of this product must R-2 (L) 500 500 500

comply with the instruction in this manual.
4. For automatic analyzers: follow their instructions.
3. MATERIALS REQUIRED BUT NOT INCLUDED
- Saline 0.9 % and high grade purified water
- Micropipet and other basic laboratory equipment.
- Calibrators and Controls (separatedly sold)
Mix well; incubate at 37 C for 1.5 minutes; measure
absorbance at 340/700 nm every 1 minute until 10 minute.
Note: See sample preparation for details of specimen
8. CALCULATION & UNIT CONVERSION
Calculation
- Calculate ∆Abs of specimen & standards vs blank

4. REAGENT COMPOSITION & PREPARATION - Plot a calibration curve NAG (U/L) = f(∆Abs)

- Reagent R-1: Citric acid solution - Calculate NAG concentration in specimen using the

- Reagent R-2: 6-methyl-2-pyridyl-N-acetyl-1-thio- curve (doing same procedure for Controls)

β-D-glucosaminide Unit conversion

- Calibrators & Controls (separatedly sold): Ready for use. 1 U/L = 0.017 µkat/L

5. SAMPLE PREPARATION & STORAGE 9. PERFORMANCE & CORRELATION TEST

- Collect urine samples without using preservatives and Performance
analyze as soon as possible. - Sensitivity: Change in absorbance when using purified

- Storing at 2 ~ 10 C, NAG in urine is stable for 1 week; water is less than 0.020; change in absorbance using NAG

storing at -20 C, NAG in urine is stable for 1 month.
- If urinary pH is more than 8.0 or less than 4.0, NAG is
inactivated; therefore, consider to environment around
storing place.
6. MEASUREMENT PRINCIPLE
This method uses 6-methyl-2-pyridyl-N-acetyl-1-thio-
β-D-glucosaminide (MPT-NAG) as a substrate. This
substrate is hydrolyzed by N-acetyl-β-D-glucosaminidase
(NAG) in urine and liberate 6-methyl-2-pyridinethiol (MPT).
The NAG activity in the sample is determined by measuring
the increase rate of the absorbance of this MPT at the
measurement wavelength of 340 nm.
solution (50 IU/L) as sample is 0.020 ~ 0.050 Abs /min.
- Specificity: The accuracy is within ±10.0%.
- Reproducibility: CV value < 5.0%.
- Measuring range: 0.5 ~ 350 U/L.
(For specimens showing a high value exceeding the upper
limit of the measurement range or specimens predicted
high in advance, dilute the specimens using physiological
saline solution).
Correlation test
Same principle, same substrate (n=52)
Regression equation Y = 1.0132X + 0.1479
Correlation coefficient R = 0.999
Same principle, different substrate (n=52)

1/2 Revision 01/2017

 Regression equation Y = 1.0175X + 0.0976 1. All specimens, as well as all instruments (e.g. test tubes)
Correlation coefficient R = 0.998 that come in contact with the specimens, must be treated by
(Y: value obtained from using UMA reagent) the following methods, or they must be treated according to
the manual for infectious medical waste provided in each
10. EXPECTED VALUES facility.
- 0.7 ~ 11.2 IU/L ・ Sterilize with an autoclave, subjecting them to high
Reference range should be established at each facility and pressure saturated steam at 121 °C for more than 20
judgement should base on measurement results in a minutes. Do not process waste containing sodium
comprehensive manner together with clinical symptoms hypochlorite solution with an autoclave.
and other measurement results. ・Immerse at least one hour in sodium hypochlorite solution
11. INTERFERENCES (active chloride concentration of over 1000 ppm).
- Ascorbic acid of 500 mg/dL, free bilirubin of 30 mg/dL, 2. This reagent contains sodium azide. Sodium azide can
albumin up to 400 mg/dL or hemoglobin of 500 mg/dL do react with lead pipe and/or steel pipe and can generate
not affect to measuring result. explosive metal azide. Make sure to use plenty of water at
disposal. Concentration of sodium azide in R-2 is 0.05%.
12. INFORMATION FOR AUTOANALYZERS
Calculation Method Rate 14. OTHER INSTRUCTIONS AND CAUTION
- Results may differ depending on the sample/reagent
Temperature 37 C
ratio. Adjust parameters for different analyzer.
Specimen 13
- Prepare the calibration curve on the day of
Volume (μL) R1 150
determination.
R2 50
Main 340
Wavelength (nm)
Sub- 700
Point 1 10
Measurement
Point 2 22
(cycle) Point 3 34

Calibration type Linear

Unit U/L

13. HANDLING, USAGE & DISPOSAL

Handling
1. Specimen can be potentially positive for infectious agents
including hepatitis B virus and HIV. Wear glove and goggle
when needed.
2. In case reagents got into skin, eye or mouth by mistake,
wash it immediately with plenty of water and consult the
doctor if needed.
3. If reagents are spilled, dilute with water and wipe it out. If
specimen is spilled, spray 80% of alcohol over the
specimen and wipe it out.
Usage
1. Store reagents under specified condition. Do not use
after expiration date.
2. Do not use the container and auxiliaries included in this
kit for other purposes.
3. Do not mix reagents of different lot for use.
4. Do not add to the reagent being used even if it is the
same lot number.
Disposal

2/2 Revision 01/2017