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ASLO direct latex express test (ASO Direct Latex)
Rapid test for semi-quantitative determination of antistreptolysin O in serum and plasma. Sensitivity >200 IU/ml. Controls positive and negative, substrate, stirrers. Analysis time no more than 3 min.
Antistreptolysin "O" (ASLO) antibodies are produced by the body during infection with beta-hemolytic streptococci in response to streptolysin "O" released from bacteria. detection and quantification ASLO in serum allows you to determine the fact and degree of infection.
Purpose and principle of the method
ASLO latex reagent is a suspension of latex particles sensitized with streptolysin "O". When the reagent is mixed with a serum sample containing ASLO antibodies, an agglutination reaction occurs, which is easily recorded visually.
Compound
- Latex reagent, 5 ml (Latex particles sensitized with streptolysin O, the reagent contains sodium azide 0.095%)
- Positive control, 0.5 ml (Human serum, positive for ASLO antibodies (>200 IU/ml), reagent contains sodium azide 0.095%)
- Negative control, 0.5 ml (Human blood serum, negative for ASLO antibodies (<200 МЕ/мл), реагент содержит азид натрия 0,095%)
- glass substrate
- Disposable stirrers
- Instruction
Test samples
ANTISTREPTOLYSIN O (ASLO)
PACKAGE
E40062 40 tests Complete set
E40060 100 tests ASLO latex reagent
E40063 100 tests Complete set
E40037 100 ml Buffer Glycine NaCl
PRINCIPLEMETHOD
Latex slide test for the qualitative and semi-quantitative determination of antistreptolysin O in undiluted serum.
METHOD
The ASLO latex test kit contains polystyrene latex particles coated with stabilized streptolysin O as an antigen that reacts immunologically with the corresponding anti-streptolysin O (ASLO) antibodies from a patient sample or control serum.
A positive reaction is determined visually by the agglutination of latex particles in the test cells on the slide.
REAGENTS
40
or
100
testsASLO latex reagent
Suspension of polystyrene latex particles coated with stabilized streptolysin O 1.0%
0.5 ml or 1.0 ml Positive control
G 
ready-to-use human serum containing the concentration of ASLO required for agglutination
0.5 ml or 1.0 ml Negative control
Ready-to-use control that does not react with latex reagent
FROM 
lay on 6 cells
buffer glycine-NaCl100 ml pH 8.2 ± 0.2
Glycine 100 mmol/l
STABILITY
Latex Reagent, Positive Control and Negative Control are stable to the stated expiration date at 2-8°C.
Do not freeze!
Damage to reagents: Presence of particles and turbidity.
SAMPLE
Serum
Stability: up to 7 days at 2-8°C,
Up to 3 months at -20°C.
Do not use highly hemolyzed or lipemic specimens.
DIGGING SCHEME
Qualitative determination (screening test)
|
Place the latex reagent, positive control, negative control, and serum samples in place at room temperature. Mix the latex particles thoroughly. |
|
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Drop by drop into individual cells on the slide: |
|
|
Positive control |
|
|
Negative control |
|
|
Latex reagent to all samples and control wells |
1 drop |
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Stir with different sticks and spread the liquid over the entire surface in certain cells. |
|
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Rotate the slide for 2 minutes so that the mixture moves slowly inside the cells on the slide, or place on an automatic rotator at 100 rpm. |
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After 2 minutes, read the results under bright artificial light. |
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(1 drop 40µl)
INTERPRETATION OF THE RESULTS
Agglutination indicates that ASLO contains more than 200 IU/ml in an undiluted sample. Serum with a positive screening test should be tested again by titration. (see part 2)
Semi-quantitative test
Samples diluted with glycine-NaCl buffer, (code E40037):
|
Breeding |
ASLO(IU/ml in undiluted sample) |
|
1
+ 1 (1: 2) |
400 |
|
Continue the test as described in part 1. |
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INTERPRETATION OF THE RESULTS
Read the titer at the final dilution step with agglutination and multiply the titer by a factor of 200 (see "Sensitivity") to get the results in IU/ml; for example: titer 1: 5 concentration of ASLO
5 x 200 [IU/ml] = 1000 [IU/ml]
CALIBRATION
The sensitivity of ASLO latex is calibrated against the international calibrator ASLO (WHO).
NORMAL VALUES
Adults: up to 200 IU/ml; Children
Each laboratory sets its own normal values.
CONTROLQUALITIES
Positive and negative controls are used with each lot. Their results should be compared with unknown samples to determine the difference between possible granulation and agglutination.
Positive control - determined by agglutination in 2 minutes.
Negative control - uniform suspension without agglutination after 2 minutes.
PERFORMANCE CHARACTERISTICS
Analytical sensitivity: -200 (±50 IU/ml)
Diagnostic sensitivity: 98%
Diagnostic specificity: 97%
Prozone effect: not seen up to 1500 IU/mL
Interference: Not observed with hemoglobin up to 10g/l, bilirubin up to 20mg/dl, lipemia up to 10g/l and rheumatoid factor RF up to 300IU/ml. Other substances may interfere.
DIAGNOSTICMEANING
An increase in ASLO titer may be associated with rheumatism and glomerulonephritis. An increased ASLO titer of more than 200 IU/mL indicates an acute streptococcal infection. The ASLO titer should be monitored every 2 weeks for 4-6 weeks.
NOTE
For professional use only.
For in vitro use only
Latex reagent, positive control, negative control and glycine-NaCl buffer contain 0.095% sodium azide. Do not swallow. Avoid contact with skin and mucous membranes.
Disposal of used reagents is carried out in accordance with
provisions of local law.
All human-derived reagents were tested using FDA-approved methods for hepatitis B surface antigen and HIV and were found to be negative. Despite the negative result, all reagents are considered to have the potential to cause infections.
Due to the drying effect, an incorrect positive result may appear after 2 minutes
As with all diagnostic methods, a definitive diagnosis cannot be made using the result of a single test, but must be based on the correlation of more than one test result with other clinical findings.
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Determination of the content of antistreptolysin O (ASO) is based on the interaction of ASO of the test sample with human streptolysin-O (CO) molecules immobilized on the surface of latex particles. When latex CO is mixed with blood serum containing ASO at a concentration exceeding 200 IU / ml, as a result of the reaction between CO and ASO, visually recorded agglutination of latex particles develops, which indicates a positive test reaction.
Research material:
Serum. The use of hemolyzed and lipemic samples is unacceptable. Samples containing fibrin particles must be centrifuged before analysis. The shelf life of the test samples at a temperature of 2-8°C is not more than 2 days.
Kit contents and storage conditions:
- CO Latex
- Buffer
- Positive calibrator - ASO concentration > 200 IU/ml
- Border calibrator - ASO concentration ~ 200 IU/ml
- Negative calibrator - ASO concentration< 200 МЕ/мл
- Test plate and spatulas.
The reagents are stable at a storage temperature of 2-8°C for 18 months. Date of manufacture, expiration date, series and catalog number are indicated on the packaging of the kit.
Preparation of reagents for the analysis procedure:
The optimal temperature of the reagents and environment during the analysis 22-25°C. At temperatures below 18°C, false results may be obtained.
CO-latex should be mixed by gentle shaking before use.
Analysis:
I. Qualitative definition:
Place the calibrators and test serum samples in a volume of 20 µl into individual wells of the test plate. Add 20 µl of CO latex to the inside of each well. Quickly mix the contents of the drops of the test sample and CO latex in each well with an individual spatula until homogeneous.
Stir the reaction mixture in a circular motion of the test plate on a mechanical orbital mixer or by hand at a speed of 80-100 rpm for 2 minutes. The agglutination process should be recorded within 1 minute after the end of the movement of the test plate. A significant increase in the time interval between stopping the test plate and fixing the result can lead to the registration of false agglutination that occurs during the drying of the reaction mixture.
Interpretation of results:
- Clearly visible aggregates of latex particles - ACO concentration > 200 IU/ml.
- Small aggregates - ASO concentration ~ 200 IU/ml.
- Uniformly homogeneous milk suspension - ASO concentration< 200 МЕ/мл.
II. Semiquantitative definition:
Prepare dilutions of test samples using buffer according to the table
Table
Analyze the resulting dilutions in accordance with paragraph I.
Reference limits :
- up to 200 IU / ml.
Specificity:
The antigen used in the CO reagent kit is highly specific for human ASO and does not cross-react with other serum proteins under testing conditions.
ATTENTION!
Do not allow the kit to freeze during storage, as this leads to the loss of the ability of the CO latex to react with agglutination, and therefore to obtaining false negative results.
Diagnostic Information :
High ASO titers indicate an acute streptococcal infection, so in this case the test must be repeated a week later to reveal the dynamics of the disease. Elevated levels of ASO in blood serum are typical for patients suffering from scarlet fever, acute rheumatoid arthritis, tonsillitis and other streptococcal infections, as well as for clinically healthy bacteria carriers.
Literature:
- Plotz and Singer, Amer. J. Med., 1979, vol. 22.
Attention!
To disinfect the slide plate, disinfectants based on free chlorine should be used - such as Chloramix, DP-2T, Javel Solid, Chlorapin.
It is unacceptable to use products based on hydrogen peroxide and fluorine, in particular "Absolucid oxy".
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