T h e C r e a t i v e A p p r o a c h t o B i o s c i e n c e

Antistreptolysin O (ASO)
Immuno-Turbidimetry
REF: 596 001 100 test
R1 Buffer 2 x 20 ml
R2 Latex 1 x 6 ml
Standard 1 x 0.2 ml

Intended Use ASO Standard:

The Standard is stable to the expiration date on the vial label when
In vitro diagnostic reagents for the quantitative determination of capped and stored at (2 - 8 ºC).
Once opened the standard is stable for 6 weeks if stored tightly
Antistreptolysin O (ASO) in human serum by means of particle- closed at 2 - 8 ºC after use.
enhanced turbidimetric immunoassay.
Specimen Collection and Preparation
Background
Serum specimens should be collected by venipuncture following
Immunological testing for specific antibodies to streptococcal good laboratory practices. Suitable assay specimens are human
metabolites provides important information regarding a prior serum samples, as fresh as possible (stored up to 2 days at 2 - 8
streptococcal infection. Antibodies are formed against both the ºC) or deep-frozen. Any additional clotting or precipitation which
pathogen itself and its metabolic products. An example for the latter occurs due to the freeze/thaw cycle should be removed by
is the antibody against streptolysin O, an enzyme secreted by centrifugation prior to assay.
betahaemolytic streptococci of the Landfield Group A. Heavily lipemic sera may lead to a non-specific reaction due to
Antistreptolysin O (ASO) testing is thus used for the diagnosis of chylomicrons. Lipemic specimens, or turbid frozen specimens after
non suppurative complications of the infections caused by these thawing, must be clarified before the assay by high-speed
pathogens: acute rheumatic fever or acute poststreptococcal centrifugation (15 min at approx. 15000 rpm).
glomerulonephritis. In the determination of antibodies to various
streptococcal exoenzymes preference is to be given to anti-streptolysin Procedure
O, since this sensitive parameter is found to be elevated in about 80
to 85% of cases. 1 - Bring the reagents and the photometer to 37ºC.
2 - Assay conditions:
Test Principle Wavelength 580 nm
Temperature 37ºC
This ASO test is based upon the ASO antigen-antibody reaction. Cuvette 1cm light path

Reagents 3 - Adjust the instrument to zero with distilled water .

4 - Pipette into a cuvette :
R1 Buffer
Phosphate buffered saline (pH 7.43) Standard Sample
Enhancer.
Sodium azide (0.095 g/L) Reagent (R1) 400 l 400 l
Standard 5l ---
R2 Latex reagent Sample --- 5l
Glycine Buffer (pH8.2)
ASO sensitized Latex (0.17 %)
Sodium azide 0.95 g/L. 5- Mix and incubate for 2 minutes,read absorbance (A1)

Standard Reagent (R2) 60 l 60l

ASO concentration is stated on the vial label.
After addition of R2, incubate and record 2nd reading after 5 minutes
Materials required but not provided with the kit (A2)
Controls
Calculation
Precautions and Warnings
Generate a reference curve by successive 1 : 2 dilutions of standard
For in vitro diagnostic use only. Do not pipette by mouth. Reagents in saline (At Least 4 points are recommended). Use Saline as zero
containing sodium azide must be handled with precaution. Sodium point. Determine  absorbance of the sample and each standard as
azide can form explosive azides with lead and copper plumbing. following:
Since absence of infectious agents cannot be proven, all specimens absorbance of sample = (A2 - A1) sample
and reagents obtained from human blood should always be handled absorbance of each standard = (A2 - A1) for each standard
with precaution using established good laboratory practices. Plot the calibration curve and obtain the result.
Disposal of all waste material should be in accordance with local
guidelines. Quality Control
As with other diagnostic tests, results should be interpreted considering
all other test results and the clinical situation of the patient. Control serum are recommended to monitor the perfomance of
manual and automated assay procedures. Each laboratory should
Reagent Preparation,Storage and Stability establish its own Quality Control Scheme and corrective actions if
controls do not meet the acceptable tolerances.
All reagents are supplied ready to use.
Reagents in the original vial are stable to the expiration date on the
vial label when capped and stored at (2 - 8 ºC).
Performance characteristics
ORDERING INFORMATION
Detection limit
CATALOG NO. QUANTITY
12.5 IU/mL

Precision 596 001 100 test

CV (%)
Intra-Run Inter-Run
Low 4.33 4.44
Medium 2.29 3.35
High 2.41 2.25

Interferences
No interference for :
Hemoglobin (1000 mg/dL)
Na-citrate (1000 mg/dL)
Heparin (50 mg/dL)
Bilirubin (60 mg/dL)
Triglyceride (2500 mg/dL)
EDTA (5 mg/dl)

Sensitivity
10.0 IU/mL.

Linearity

400 IU/mL.
Specimens showing higher concentration should be diluted 1+4
using physiological saline and repeat the assay (result×5).

Expected Values
Normal values 0 - 200 IU/ml
Each laboratory should establish an expected range for the geographical
area in which it is located.

References
1.Tadzynsky LA, Ryan ME. Diagnostic of rheumatoid fever. A
guide to criterial and manifestations. Postgrad Med 1986;
79:295.
2.Dillon,H.C.jr,Reeves M.A,aM.J.Med,56,333-346(1974).
3.Bach GL, Cadotte R, Wiatr RA, et al. Latex antiestreptolysin O
test as a tube dilution procedure. Am J Clin Pathol 1972; 57:
209.
4.Klein,G.C.Baker,C.N,Jones,W.L,21,999-1001(1971).
5-Curtis GDW, Kraak WAG, Mitchell RG. Comparison of latex and
hemolysis tests for determination of antiestreptolysin O (ASO)
antibodies. J Clin Pathol 1988; 41: 1331.

Egyptian Co for Biotechnology - Spectrum Diagnostics (S.A.E)

Obour city industrial area. block 20008 piece 19 A. Cairo. Egypt.
Tel: +202 4489 2248 - Fax: +202 4489 2247
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