Immuno-turbidimetric test for the quantitative determination of ASO (Anti-Streptolysin O) antibodies in human serum on Beckman Coulter analysers.
SUMMARY AND EXPLANATION
Reference1,2 Group A streptococcus is one of the most common causes of human bacterial infections, causing diseases such as acute rheumatic fever, acute glomerulonephritis, acute pharyngitis, sinusitis, pneumonia, septic scarlet fever, gangrene, and lymphangitis. Streptolysin O is a haemolysin produced by group A streptococci. In an infected individual streptolysin O acts as a protein antigen to which the patient mounts an antibody response. Titres rise as early as 1 week and peak 3 – 6 weeks after infection. In the absence of complications or re-infection, the ASO titre will usually fall to pre-infection levels within 6 – 12 months.
METHODOLOGY
When a sample is mixed with R1 buffer and R2 latex solution, Anti-Streptolysin O antibodies react specically with Streptolysin-O coated latex to yield insoluble aggregates. The absorbance of these aggregates is proportional to the ASO concentration in the sample.
SPECIMEN SPECIMEN STORAGE AND STABILITY
Serum: Stable for 8 days when stored at 2…8 °C and 2 days when stored at 15…25 °C.3
Instructions For Use BLOSR6x94 03 EN ASO
MAY 2015 Page 1 of 5 REAGENTS WARNING AND PRECAUTIONS
Exercise the normal precautions required for handling all laboratory reagents. To avoid the possible build-up of azide compounds, ush waste-pipes with water after the disposal of undiluted reagent. Dispose of all waste material in accordance with local guidelines. Safety data sheet available for professional user on request.
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.
CAUTION Sodium azide preservative may form explosive compounds in metal drain lines. See NIOSH Bulletin: Explosive Azide Hazard (8/16/76). To avoid the possible build-up of azide compounds, ush wastepipes with water after the disposal of undiluted reagent. Sodium azide disposal must be in accordance with appropriate local regulations.
GHS HAZARD CLASSIFICATION
Not classied as hazardous
REAGENT PREPARATION
R1 is ready for use and can be placed directly on board the instrument. R2 should be mixed by inversion 5 – 10 times before placing on board the instrument and at weekly intervals thereafter.
STORAGE AND STABILITY
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8 °C. Once open, reagents stored on board the instrument are stable for 60 days. Do not use if there are visible signs of microbial growth, turbidity of precipitate, or any change in reagent colour.
CALIBRATION CALIBRATOR REQUIRED
This assay may be calibrated using multi-point calibration or single point calibration assay. For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021. For single point calibration Serum Protein Multi-Calibrator Cat. No.: ODR3021, Calibrator 4 may be used. MC Cal A Cat No. ODR30037 for Mastercurve enabled systems only. Please refer to User Guide for further instructions.
ASO EN Instructions For Use BLOSR6x94 03
Page 2 of 5 MAY 2015 Calibrator ASO values are traceable to the WHO NIBSC 1st International standard for Anti- Streptolysin O, AST.4, 5 Recalibrate the assay when the following occur: Change in reagent lot or signicant shift in control values; Major preventative maintenance was performed on the analyser or a critical part was replaced. Following calibration, the resulting curve should be visually reviewed for acceptability, on the Beckman Coulter analyser, using the software options to access the Calibration Monitor. Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
QUALITY CONTROL ITA Control Sera ODC0014, ODC0015 and ODC0016 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 specic 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 specied limits.
TESTING PROCEDURE(S) Refer to the appropriate User Guide and Setting Sheet for analyser-specic assay instructions for the sample type as listed in the Intended Use statement.
CALCULATIONS The Beckman Coulter analysers automatically compute the ASO concentration of each sample.
REPORTING RESULTS REFERENCE INTERVALS
Reference1 Adults ≤ 200 IU/mL Children ≤ 150 IU/mL
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 are representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these values.
Instructions For Use BLOSR6x94 03 EN ASO
MAY 2015 Page 3 of 5 PROCEDURAL NOTES LIMITATIONS
Samples with extremely abnormal optical characteristics, especially turbidity, may produce atypical results.
INTERFERENCES
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows: Icterus: Interference less than 5% up to 40 mg/dL or 684 µmol/L bilirubin Haemolysis: Interference less than 3% up to 5 g/L haemoglobin Lipemia: Interference less than 5% up to 1000 mg/dL Intralipid®. Refer to Young6 for further information on interfering substances.
PERFORMANCE CHARACTERISTICS LINEARITY
The test is linear within a concentration range of 100 – 1000 lU/mL.
SENSITIVITY
The lowest detectable level in serum on an AU400 analyser was calculated as 7 IU/mL. The lowest detectable level represents the lowest measurable level of Anti-Streptolysin O 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 ASO OSR6194 assay on the AU2700 against another commercially available ASO method. Results of linear regression analysis were as follows: y = 1.031x - 16.575 r = 0.992 n = 102 Sample range = 106 – 941 IU/mL
PRECISION
The following data was obtained on an AU640 using 3 serum pools analysed over 20 days. n = 80 Within-run Total Mean (IU/mL) SD CV% SD CV% 152 2.50 1.65 3.77 2.49 318 3.84 1.21 7.74 2.43 644 7.61 1.18 16.90 2.63
ADDITIONAL INFORMATION # User dened ¤ Analyser default value † For multi-point calibration use Serum Protein Multi-Calibrator Cat. No.: ODR3021. For single-point calibration use Serum Protein Multi-Calibrator ODR3021 (Cal. No. 4), with calibration type “AB”. Set the factor range at 0 to 99999.9 when using calibration type “AB”. ₪ MC Cal A Cat. No. ODR30037 * Values set for working in IU/mL
ASO EN Instructions For Use BLOSR6x94 03
Page 4 of 5 MAY 2015 REFERENCES 1. Thomas L. Streptococcus pyogenes infection. In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:1201-1203.
3. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and serum samples. WHO/DIL/LAB/99.1 Rev.2:22pp.
4. Blirup-Jensen S, Johnson AM, Larsen M. Protein standardization IV: value transfer procedure for the assignment of serum protein values from a reference preparation to a target material.Clin Chem Lab Med 2001;39:1110-1122.
5. Spaun J, Bentzon MW, Olesen Larsen S, Hewitt LF. International standard for anti-streptolysin-O. Bull Wld Hlth Org 1961;24:271-79.
6. Young DS. Effects of drugs on clinical laboratory tests, 5thed. AACC Press, 2000.
