Bilirubin Direct Blosr 6x11

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OSR6111 4 x 6 mL R1 (DBILC), 4 x 6 mL R1 (DBILB) OSR6211 4 x 20 mL R1 (DBILC), 4 x 20 mL R1 (DBILB) OSR6611 4 x 173 mL R1 (DBILC), 4 x 173 mL R1 (DBILB)

For in vitro diagnostic use only.

ANNUAL REVIEW

Reviewed by Date Reviewed by Date

Photometric colour test for the quantitative determination of direct bilirubin in human serum and plasma on Beckman Coulter analysers.

OSR6611 for use on the AU5800, AU2700 and AU5400 systems only.

SUMMARY AND EXPLANATION

Reference1,

80 – 85% of bilirubin produced daily originates from haemoglobin released by the breakdown of senescent erythrocytes, the remaining 15 – 20% results from the breakdown of haem-containing proteins such as myoglobin, cytochromes, catalases and from bone marrow as a result of ineffective erythropoiesis.

Because of its poor solubility in water unconjugated bilirubin (indirect bilirubin) is transferred to the liver bound to albumin. Inside the hepatocytes it is rapidly conjugated with glucuronic acid to produce bilirubin mono- and diglucuronide (direct bilirubin) which are then excreted in bile together with all other normal biliary constituents.

Whereas prehepatic jaundice (e. haemolytic anemia and neonatal jaundice) is primarily associated with an increase in unconjugated bilirubin, the assessment of direct bilirubin is helpful in the determination of hepatic and post-hepatic jaundice. Diseases of hepatic origin with predominantly conjugated hyperbilirubinemia include acute and chronic viral hepatitis, liver cirrhosis and hepatocellular carcinoma. Diseases of post hepatic origin with predominantly conjugated hyperbilirubinemia include extrahepatic cholestasis and liver transplant rejection. Chronic congenital conjugated hyperbilirubinemias include Dubin-Johnson and Rotor syndrome. The differentiation between chronic congenital hyperbilirubinemias and acquired types of bilirubininemia is accomplished via the measurement of bilirubin fractions and the detection of normal liver enzyme activities.

METHODOLOGY

Reference 3

Instructions For Use BLOSR6X11 06 EN DBili Metabolite JANUARY 2016 Page 1 of 6

A stabilised diazonium salt, 3,5 Dichlorophenyl diazonium tetrauoroborate (DPD) couples directly with direct (conjugated) bilirubin in an acid medium to form azobilirubin. The absorbance at 570 nm is proportional to the direct bilirubin concentration in the sample.

CHEMICAL REACTION SCHEME

Bilirubin + DPD Azobilirubin

Serum and heparinised plasma: stable for 3 days when protected from light and stored 15..°C. 1

Even slight haemolysis can cause a reduction in value and such samples should be avoided.

Lipemic samples should also be avoided.

Hazard Warnings and Risk Phrases:

DBILB and DBILC: Corrosive, Contains sulphuric acid. R35; Causes severe burns.

Safety Phrases:

S20, S26, S30, S36/37/39, S45, S60. When using do not eat or drink. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Never add water to this product. Wear suitable protective clothing, gloves and eye/face protection. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible). This material and its container must be disposed of as hazardous waste.

Dispose of all waste material in accordance with local guidelines.

Refer to Safety Data Sheets for further information.

REACTIVE INGREDIENTS

Final concentration of reactive ingredients:

3,5 Dichlorophenyl diazonium tetrauoroborate 0 mmol/L

The concentrations of the reactive components of the reagents shown on the kit label are the actual concentrations in the individual R1/R2 vials. The reagent composition which is shown in the Instructions For Use is the nal concentration of these components in the reaction cuvette after addition of R1, Sample, and R2.

GHS HAZARD CLASSIFICATION

Direct Bilirubin Blank R1 DANGER

H314 Causes severe skin burns and eye damage. P280 Wear protective gloves, protective clothing and eye/face protection.

DBili Metabolite EN Instructions For Use BLOSR6X11 06 Page 2 of 6 JANUARY 2016

Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.

Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient samples are tested and each time calibration is performed.

The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory generates analyte specic control target values and intervals based on multiple runs according to their requirements. These target values should fall within the corresponding acceptable ranges given in the relevant product literature.

If any trends or sudden shifts in values are detected, review all operating parameters.

Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specied limits.

Refer to the appropriate User Guide and Setting Sheet for analyser-specic assay instructions for the sample type as listed in the Intended Use statement. The paediatric application is suitable for use with small volume serum/plasma samples.

The Beckman Coulter analysers automatically compute the direct bilirubin concentration of each sample.

Reference 1 Adults and Children < 3 μmol/L (< 0 mg/dL)

Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results should always be assessed in conjunction with the patient’s medical history, clinical examinations and other ndings.

Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these values.

Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:

Lipemia: Interference less than 10% up to 300 mg/dL Intralipid®

In very rare cases gammopathy, especially monoclonal IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

Refer to Young 4 for further information on interfering substances.

DBili Metabolite EN Instructions For Use BLOSR6X11 06 Page 4 of 6 JANUARY 2016

The test is linear within a concentration range of 0 – 171 μmol/L (0 – 10 mg/dL).

SENSITIVITY

The lowest detectable level on an AU600 analyser was calculated as 0 μmol/L.

The lowest detectable level represents the lowest measurable level of direct bilirubin that can be distinguished from zero. It is calculated as the absolute mean plus three standard deviations of 20 replicates of an analyte free sample.

METHODS COMPARISON

Patient serum samples were used to compare this Direct Bilirubin (OSR6111) assay on the AU640 against another commercially available direct bilirubin assay. Results of linear regression analysis were as follows:

y = 0 – 0 r = 0 n = 99 Sample range = 0 – 151 μmol/L

Estimates of precision, based on CLSI recommendations 5 , are consistent with typical performance. The within run precision is ≤5% CV or SD ≤ 2 μmol/L and total precision is ≤7% CV or SD ≤ 3 μmol/L. Assays of control sera and plasma were performed and this data reduced following CLSI guidelines above.

The following data was obtained on an AU2700 using 3 plasma pools analysed over 20 days.

n = 80 Within-run Total Mean μmol/L SD CV% SD CV% 13 0 2 0 3. 22 1 4 1 5. 106 4 3 4 4.

‡ The above parameters must be entered twice using test names DBILC (colour) and DBILB (blank). Set the test as SAMPLE BLANK in the INTER RELATED TEST menu

¤ Analyser default value † System Calibrator Cat. No.: 66300

§ Set the factor range for the blank reagent at –99999 to 99999

ж Set the test as SAMPLE BLANK in the COMMON TEST PARAMETERS TEST NAME SAMPLE BLANK menu.

Instructions For Use BLOSR6X11 06 EN DBili Metabolite JANUARY 2016 Page 5 of 6