Athenese DX LDH -P Biochemistry Test Kit

INTENDED USE The TRUEchemie LDH-P test kit is intended to use for the in vitro quantitative kinetic determination of Lactate Dehydrogenase activity in serum.   INTRODUCTION The enzyme lactate dehydrogenase (LDH) is distributed in tissues particularly heart, liver, muscle, and kidney. The enzyme found in circulation is a mixture of five isoenzymes based on their mobility. Elevated serum levels of LDH are found in serum in myocardial infarction, liver disease, renal disease, certain forms of anemia, malignant diseases, and progressive muscle dystrophy. LDH catalyzes the oxidation of pyruvate to lactate accompanied by the simultaneous reduction of NADH to NAD.LDH activity in serum is proportional to the decrease in absorbance due to reduction of NADH   REAGENTS COMPOSITION Working reagent composition Tris Buffer :90 mmol/L Sodium Pyruvate :1.20 mmol/L Sodium chloride :150 mmol/L NADH :0.24 mmol/L   STORAGE AND STABILITY The components of the kit, stored at 2 - 8^oC, will remain stable until the expiry date stated on the label.   REAGENT PREPARATION The working reagent is prepared by mixing four (4) volumes of R1 with one (1) volume of R2 in a disposable container. Example: 20ml R1 5ml R2 The working reagent is stable for 30 days at 2-8°C   SAMPLE / SPECIMEN AND STORAGE Serum without hemolysis. Samples should be assayed soon after collection. LDH in serum is reported to be stable for 2-3 days at room temperature. Liver LDH is particularly labile and is destroyed if frozen or thawed.   WARNINGS AND PRECAUTIONS

1. For in vitro diagnostic use.
2. Specimens should be considered infectious and handled appropriately.
3. Avoid ingestion. DO NOT PIPETTE BY MOUTH.
4. The reagent contains sodium azide as a preservative. Avoid skin and eye contact.
5. Protect from direct light.
6. Avoid microbial contamination.

  MATERIALS REQUIRED BUT NOT PROVIDED

1. Pipettes to accurately measure required volumes.
2. Test tubes/rack
3. Timer
4. 37 °C heating block or water bath
5. Photometer capable of accurately measuring absorbance at 340 nm

  TEST PROCEDURE Primary wavelength 340 nm Temperature 37 °C Prewarm the reagent to reaction temperature.

  Reading: Blank the Photometer with Distilled water. Mix well, read the initial absorbance after 60 sec and repeat the absorbance reading after every 30 sec upto 90 second.   Calculations: ΔE = Initial absorbance - Absorbance after 1st/2nd /3rd reading. Calculations determine ΔE/min. for every reading Find the mean value of ΔE/min. ΔA = (Avg ΔE/min) x 8095 = U/L of LDH   NORMAL VALUES Adult : 235 - 470 IU/L at 37 °C Children : <12 years old have LDH levels 10-15% higher than adult ones It is strongly recommended that each laboratory establish its own normal range   AUTOMATED PROCEDURE Appropriate program sheet is available for different analyzers upon request.   LIMITATIONS OF TEST Linearity : 2000 U/L. Sensitivity : 3.4 U/L.   Samples that have LDH values greater than 2000 U/L should be diluted 1:1 with saline, re-assayed and the results multiplied by 2.   INTERFERENCES

1. Oxalate, oxamates, and EDTA will inhibit LDH.

Young, et. al. gave a list of drugs and other substances interfere with the determination of LDH activity.   SYSTEM PARAMETERS Mode : Kinetic Factor : 8095 Wave length : 340 nm Units : U/L Flow cell temp. : 37 °C Blank : Distilled water Working reagent volume : 1000 μL Sample volume : 20 μL Reaction. Slop : Decreasing Delay (Lag) time : 60 Sec. Read time : 90 Sec. Low normal : 235 High normal : 470