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www.biomaghreb.com
IVD
FT En 44 Date of updating : 10/ 2020 Version B
IN VITRO DIAGNOSTIC USE
Manufacturer Use by In Vitro Diagnostic Temperature
Limitation
Catalogue number See insert Keep away
from light
Sufcient
for < n > essais
Batch number
STREPTOLYSIN-O
Determination of anti streptolysin - O
PRINCIPLE
A,C,G groups of haemolytic b streptococcus secreted an enzyme : Streptolysin- O. wich present an
haemolytic activity in its reduced form. This streptolysin-O causes antibody formation revealed by a
neutralization reaction of its enzymatic haemolytic activity towards rabbit red blood cells.
REAGENT COMPOSITION
- Streptolysin lyophilisate
- Streptolysin buffer in 10 times concentrated form: It is to be diluted for use: 1 part buffer to 9 parts
distilled water, in case of crystallization, reheat to 37°C to put the crystals back into solution.
A 1% rabbit red blood cell suspension in streptolysin buffer 1x is conventionally used. However, the
high titre of the streptolysin used makes it possible to obtain identical results with very carefully
washed human red blood cells of group O: 3 washes in physiological water. The last base is taken up
in the streptolysin 1x buffer.
Reconstitution of the titrated Streptolysin: Insert a hollow needle through the rubber stopper so that
the vial is filled with air without risk of losing any of the lyophilisate.
Uncap and uncap the vial to introduce exactly 8 ml of isotonic buffered solution. Recap and promote
solution by shaking moderately. The streptolysin is then in its active form.
SAFETY CAUTIONS
Biomaghreb reagents are intended for qualified personnel, for in vitro use (do not pipette with the
mouth).
• Consult the current MSDS available on request or on www.biomaghreb.com;
• Check the integrity of the reagents before use; and
• Disposal of waste: comply with the legislation in force.
For safety reasons, treat any specimen or reagent of biological origin as potentially infectious. Res-
pect the legislation in force.
MACROMETHOD SAMPLES
The sera to be examined must not show haemolysis. They must be inactivated by heating at 56°C for
30 minutes (If sera which have been inactivated for more than 24 hours are used, it is recommended
that they be placed again in a water bath at 56°C for 10 minutes).
PRESERVATION AND STABILITY
- Lyophilisate : our streptolysin has a shelf life of about 3 years at + 4° C.
- Reconstituted: it must be used within 5 hours.
PROCEDURE
(See Table 1)
Primary dilutions :
In 2 haemolysis tubes A and B, introduce 2 ml of buffered isotonic solution, add 0.5 ml of serum
to be studied in tube A (dilution made 1/5). Mix and transfer 1 ml from tube A to tube B (dilution
carried out at 1:15).
Preparation of the titration dilutions of the sera to be examined.
For each serum, prepare 2 sets of tubes numbered 1-2-4-6-8-10 for serum stock dilution A (at 1/5)
and 3-5-7-9 for serum stock dilution B (at 1/15) and two tubes 11 and 12 for the red blood cells and
streptolysin controls.
Proceed according to Table 1 in geometric progression. Distribute the volumes indicated in ml.
Table 1 :
Usual values are less than 200 ASU samples with value equal
or greater than 200 ASU are considered pathological.
MICROMETHOD
Prepare the dilutions of the test sera (See Table 2):
- Dilution 1/5: 0.1 ml of serum + 0.4 ml of buffer
- Dilution 1/15: 0.1 ml serum + 1.4 ml buffer
Spread on microplate (in ml)
Table 2 : MICROMETHOD
Usual values are less than 200 ASU. Samples with value equal
or greater than 200 ASU are considered pathological.
