FEWF0025

5x5ml

STORE AT 2 – 8°C

ISO 13485 accredited company FOR IN-VITRO DIAGNOSTIC USE ONLY FEWF – WEIL-FELIX Page 1 of 2

FEBRILE SERODIAGNOSTICS Qualitative Test Procedure:

1. Bring the reagents and samples to room temperature.
Somatic (O) and Proteus antigens: 48 – 50°C for 4 hours
Flagellar (H) antigens: 48 – 50°C for 2 hours.
2. Place 50µl or one drop of the sample and 1 drop of each control 8. Examine for signs of agglutination. The titre to be taken is
(WEIL-FELIX) 3.
into separate circles on the card.
Resuspend the antigen gently.
the last tube to show agglutination.

4. Add one drop of the latex reagent to each circle next to the Tube No. Saline ml Serum ml Dil/Titre
AGGLUTINATION FOR SLIDE AND TUBE TESTS sample which is to be tested.
5. Mix with the disposable pipette / stirrer and spread over the 1 1.9 ml 0.1 ml 1/20
Principle: entire area enclosed by the ring. Use a new stirrer for each 2 1.0 ml 1/40
Febrile Antigens are standardised suspensions of stained bacteria sample.
3 1.0 ml 1 ml 1/80
prepared for the rapid detection and semi-quantitation of serum 6. Rotate the cards at 100 r.p.m. for 2 minutes.
antibodies developed during the acute stage of the disease. The 4 1.0 ml Serial 1/160
antigens agglutinate in the presence of the homologous antibodies in Circle 1 80µl 5 1.0 ml Dilution 1/320
the sample tested.
Circle 2 40µl 6 1.0 ml 1/640
Presentation: Circle 3 20µl 7 1.0 ml 1/1280

Contents Code Circle 4 10µl 8 1.0 ml Blank

Circle 5 5µl Discard 1 ml
Brucella Abortus FEBBAB05 5ml
Brucella Melitensis FEBBME05 5ml 1. Add 1 drop of Febrile Antigen Suspension to each circle. NOTE: IF USING A WATERBATH IT IS VERY IMPORTANT TO ENSURE THAT
Proteus OXK FEPOXK05 5ml 2. Mix well using a pipette / stirrer. THE WATER REACHES AT LEAST TWO THIRDS OF THE WAY UP THE TUBE TO
3. Rotate the slide by hand or on a mechanical rotator at 100 r.p.m. for 2 ENSURE THAT THE CONVECTION CURRENTS WITHINTHE TUBE ARE
Proteus OX2 FEPOX205 5ml MAINTAINED THUS ELIMINATING THE POSSIBILITY OF FALSE RESULTS.
minutes.
Proteus OX19 FEPOX905 5ml 4. Agglutination in any of the circles is indicative of the following results:
5. Quality Control:
Composition: Each run of tests should be validated with a positive and negative
80µl 1:20
Salmonella Febrile Antigens Blue stained Antigens specific control.
to somatic ‘O’ antigens. 40µl 1:40
Red stained Antigens specific 20µl 1:80 Reading And Interpretation:
To somatic flagellar ‘H’ antigens • Examine macroscopically for the presence or absence of
Positive Control Human Serum 10µl 1:160 clumps or agglutination within 1 minute of removing the card
Sodium Azide 0.95g/L. 5µl 1:320 from the rotator, comparing the results with the controls.
• Negative results show no signs of agglutination.
Negative Control Animal Serum
Sodium Azide 0.95g/L. Tube Agglutination Test:
1. Label 8 small plastic tubes as set out in the table below. Limitations Of The Procedure:
2. Make a 1/20 dilution of serum and saline in the first tube. • False negative results can be obtained in the early stages of the
Storage:
3. Take 1 ml of the 1/20 dilution in Tube 1, transfer to Tube 2 and disease as well as in cases of immunounresponsiveness and
Store components at 2-8°C.
proceed to make serial dilutions as shown below until Tube 7. antibiotic treatment.
Discard 1ml from Tube 7. • False negative somatic (O) results may occur in typhoid patients
Samples:
4. Tube 8 serves as a blank or control containing only saline. who have been treated with chloramphenicol.
• Serum stable for 7 days at 2-8°C,
5. Dilute Positive and Negative Controls 1/10 with 9g/L saline. • The sensitivity of the test may be reduced at low temperatures.
• Samples should be free from contamination, haemolysis and
6. Add one drop of the appropriate antigen suspension to each The best results are achieved at +10°C.
Lipemia.
tube and mix well. • In some geographical areas with a high prevalence of febrile
7. Incubate as follows:- Antigens at 36°C for 24 hours. antibodies, it is recommended that the sample is diluted ¼ in
Additional Equipment:
The incubation process may be accelerated by incubating as 9g/L saline.
Glass Slides and a Mechanical Rotator set at 100 r.p.m.
follows:

Fortress Diagnostics Limited, Unit 2C Antrim Technology Park, Antrim BT41 1QS (United Kingdom) REVISED OCT/07
TEL: +44 (0) 2894 487676 FAX: +44 (0) 2894 469933 www.Fortressdiagnostics.com
 FEWF0025

5x5ml

STORE AT 2 – 8°C

ISO 13485 accredited company FOR IN-VITRO DIAGNOSTIC USE ONLY FEWF – WEIL-FELIX Page 2 of 2

Notes:
1. The sensitivity of the test may be reduced at low temperatures.
Optimum results are achieved over 10°C.
2. When testing for Brucella Antibodies it is recommended to
reduce sample volume to 0.02ml in order to avoid prozone.
3. In some geographical areas with a high prevalence of febrile
antibodies, it is recommended that the sample be diluted 1 / 4
with NaCl 9 g/L prior to testing.
4. Controls are designed to assess what results should be achieved
in both positive and negative results.
5. Delay in reading the results may result in false positive reactions.
6. The incubation times may be accelerating by incubating as
follows:
7. Somatic (O) and Proteus Antigens: 48 – 50°C for 4 hours
8. Flagellar Antigens: 48 – 50°C for 2 hours
9. A somatic (O) reaction is characterised by coarse, compact
agglutination which tends to be difficult to disperse, while the
flagellar (H) has a characteristic loose, flocculant agglutination.

Reference:
Felix A. Trans Rpy Soc Trop Med Hyg 1944; 37: 321 – 325
Weil E. and Felix A. Wein Klin. 29:974 (1916)
Gualtney JB et al. Appli Micro 1971; 22: 635 – 640

For In Vitro Diagnostics Use Only

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Warning, Read Enclosed Documents

Instructions For Use

Manufactured By

Fortress Diagnostics Limited, Unit 2C Antrim Technology Park, Antrim BT41 1QS (United Kingdom) REVISED OCT/07
TEL: +44 (0) 2894 487676 FAX: +44 (0) 2894 469933 www.Fortressdiagnostics.com