Instructions for use

Intended use
TUBEX® TF is a semi-quantitative in vitro diagnostic serum assay for the detection of acute
typhoid fever infection caused by Salmonella typhi.

Principle of the assay

TUBEX® TF detects the presence of anti-O9 antibodies in the patient’s serum by assessing
their ability to inhibit the reaction between the antigen coated brown and antibody coated
blue reagents. The level of inhibition is proportional to the concentration of anti- O9
antibodies in the sample. Brown reagent contains iron and separation is enabled by
magnetic force. The result is read visually against a color scale. A positive TUBEX® TF result,
together with symptoms of typhoid fever, is a strong indication of a typhoid infection.

Assay specificity
TUBEX® TF specifically detects IgM antibodies to the S. typhi O9 lipopolysaccharide antigen.
This antigen is highly specific to S. typhi and other Salmonella serogroup D bacteria by its
extremely rare sugar (α-D-tyvelose). IgM anti-O9 antibodies are normally not present in
healthy individuals.

Specimen collection and storage

Use clear serum samples or heparin plasma. Do not use EDTA or citrate plasma. Avoid
grossly lipemic, icteric or hemolysed samples. Colored patients samples, e.g. by hemoglobin
(read color) and bilirubin (green color), may disturb the color of the supernatant and result
in indeterminate assay results. Colored samples may be analyzed by using TUBEX® TF wash
kit (REF o 10-501. The serum samples should be stored at 2-8 0C or frozen (≤-18 0C), if not
used immediately.
Precautions for users
1. TUBEX® TF is for in vitro diagnostic use only.
2. Reading of results requires a normal color vision.
3. Do not use the kit after expire date.
4. Do not mix reagents from different lots or kits.
5. Avoid microbiological contamination of reagents.
6. Caution! Strong magnets inside the TUBEX® TF color scale.
7. Wear protective gloves and protective goggles.
8. All patient specimens should be regarded as contagious and handled and disposed of
according to appropriate regulations.
9. If reagents come in contact with eyes, flush with plenty of water and seek medical
advice.
10. ProClin 300 (60 ppm) used as a preservation in this product might be allergenic.
11. Safety data sheets are available on request.

Components in TUBEX® TF
Materials supplied for 36 determinations.
TUBEX® TF Brown Reagent; 1 vial, 1.7 ml, antigen coated magnet particles in protein
stabilized buffer, pH 8.2. preservative added. Ready for use.
TUBEX® TF Blue Reagent; 1 vial, 3.5 ml, antibody coated particles in protein stabilized
buffer, pH 8.2. preservative added. Ready for use.
TUBEX® TF Negative Control; 1 vial, 0.4 ml, protein stabilized buffer, pH 8.2. Yellow color,
preservative added, ready for use.
TUBEX® TF positive control; 1 vial, 0.4 ml, control antibody in protein stabilized buffer, pH
8.2. preservative added, ready for use.
TUBEX® TF reaction well strip; 6 strips (6 reaction wells per strip single-use.
TUBEX® TF sealing tape; 10 sealing tapes for reaction wells. Single-use.
TUBEX® TF certificate; certificate of lot content.
TUBEX® TF reaction well strips and TUBEX® sealing tapes are for single use only due to risk
of contamination and false positive result. Additional TUBEX® reaction well strips are
available (REF No 10-929).
Material required but not provided
(not provided in the test kit but can be ordered separately, please contact your respective
distributor)
TUBEX® Color scale (REF 10-928), required for separation and scoring of results, reusable.
The magnet in the TUBEX® color scale is permanent. Precision pipettes, shaker, vortex and
timer.
Additional materials and special equipment
(not provided in the test kit but can be ordered separately, please contact your respective
distributor).
Stickers for color scale (REF 91-336). Our recommendation is to add a news sticker on the
color scale every 6 months.
Schematic assay procedure

Assay procedure
Carefully read the instructions for use before starting the analysis (also see assay procedure
above). The procedure is also described in an instructional film available on IDL biotech AB
home page www.idl.se.
The assay should be performed at room temperature, (18-28 0C). inclusion of controls are
recommended at every test occasion, but not required for every reaction well strip used.
Preparations
Place the TUBEX® reaction well strips on the bench, for the well to reach room temperature.
Color label each component’s bottle and cap (optional). Vortex or shake all reagents and
samples thoroughly prior to use.
Check visually that all sediments have dispersed into solution.
Procedure
The accuracy of the test is related to adherence to the assay procedure and accurate
volume pipetting.
1. Place the TUBEX® reaction well strip upright on the bench, well number face-front (d not
place the strip on the color scale yet).
Add 45 µl sample, TUBEX® TF Brown reagent to each well.
2. Add 45 µl sample, TUBEX® TF positive control or TUBEX® TF negative control to
appropriate wells, and carefully mix by pipetting up and down 5-10 times. Thorough
mixing is essential. Avoid foaming. Use a new pipette tip for each sample.
3. Incubate on the bench for 2 minutes.
4. Add 90 µl TUBEX® TF blue reagent to each well. Be careful not to touch the reaction
mixture in the well.
5. Cover the TUBEX® reaction well strip with a TUBEX® sealing tape. (make sure there is no
condense/or liquid on the strip surface). Press the tape hard against the plastic to
prevent leakage.
Mix thoroughly for 2 minutes using the following procedure :
- Hold the TUBEX® reaction well strip at one end using one hand’s thumb and pointing
finger.
- Tilt the TUBEX® reaction well strip horizontally (900) to expose maximum well surface
for the mixture.
- Shake the TUBEX® reaction well strip rapidly backwards and forwards for 2 minutes.
Make sure that the contest flow across the entire exposed well surface.
6. Place the TUBEX® reaction well strip on the TUBEX® color scale as far to the left as
possible. To obtain a clear supernatant, allow separation for 5 min, then read and
interpret the result.
Interpretation of Results
Read and score the results by comparing the color of each supernatant to the TUBEX® color
scale. Reading should be performed at good light conditions after 5 minutes, although
results are stable for at least 30 minutes if not disturbed. Do not interpret results or re-
separate reactions if the well strip is accidently tipped over during or after separations.
The scores range from 0 (clear pink) to 10 (intense blue).
A general guide to the interpretation of TUBEX® TF scores is presented below.
Interpretation should focus on the detection of any blue tone in the supernatant,
particularly important regarding the lower scores.
To indicate a correct performance of the kit. TUBEX® TF negative control should score ≤ 2
and TUBEX® TF positive control should score ≥ 8.
Assay interpretation guide
Score Result Interpretation guide
0-2 Negative Does not indicate current
typhoid fever infection.
TUBEX® TF negative control
>2 or <4 Inconclusive Repeat the test. If still
inconclusive, repeat
sampling at a later date
4-10 Positive The higher score the
stronger is the indication of
current typhoid fever
infection
TUBEX® TF positive control
Indeterminate
No clear score obtained due to :
1. Poor adherence to assay protocol
2. Poor specimen quality (hemolysis, heavily icteric)
Recommendations :
1. Retest or repeat sampling and retest
2. Use TUBEX® TF wash (REF 10-501) according to procedure
and retest.

If TUBEX® TF indicates current typhoid fever but blood culture is negative, the following may
be the reasons :
- Patient truly has typhoid fever. Culture is commonly positive in only 50-80% of affected
patients.
- Concomitant antibiotic intake cloud give false negative blood culture result and a
positive TUBEX® TF test result.
Reagents Storage
The kit should be stored at 2-80C. do not freezel
Store the reagents upright in their original container if not used at once at once.
Assay characteristic
TUBEX® TF has an analytic sensitivity of 15-20 µg/ml of O9 specific IgM antibodies, as
measured with monoclonal antibodies.
Limitations of the procedure
TUBEX® TF results should be interpreted in conjunction with all available clinical
information.
Although TUBEX® TF specifically detects acute phase antibodies (IgM), it may also detect
convalescent phase antibodies (IgG). IgG antibodies, not reactive by themselves, can
synergistically bind to the antigen coated particles.
TUBEX® TF may not detect cases where IgM antibodies are present in extremely low levels,
such as early in the infection when the immune system has not yet been sufficiently
stimulated.
Accessories
Product REF
TUBEX® color scale 10-928
TUBEX® color scale sticker 91-336
TUBEX® well strips package 10-929
TUBEX® workstation 10-930
TUBEX® pipett 45 ul 10-950
TUBEX® pipett 90 ul 10-951
TUMIX shaker 10-931
All accessories and additional material can be ordered at [email protected]
Warranty
The performance data presented here were obtained using the procedure indicated. Any
change or modification in the procedure, not recommended by IDL Biotech AB, may affect
the results. In such event, IDL Biotech AB disclaims all warranties expressed, implied or
statutory, including the implied warranty of merchantability and the fitness for use.

References
Kawano RL et al. comparison of serological test kits for diagnosis of typhoid fever in the Lim,
Philippines. J Clin Microblol 2007; 45:246-247.
Lim, PL et al, One-step 2-minutes test to detect typhoid-specific antibodies based on particle
separations in tubes. J Clin Microblol 1998; 36:2271-2278.
Olsen, SJ et al. evaluation of rapid diagnostic tests for typhoid fever. J Clin Microbiol 2004;
42; 1885-1889.
Tarupiwa A et al. evaluation of TUBEX TF and OnSite Typhoid IiG/IgM Combo rapid tests to
detect Salmonella enterice serovar typhi infection during a typhoid outbreak in Harare,
Zimbabwe. BMC researches notes (2015) 8:50
Tam, FCH et al. the TUBEX typhoid test based on particle-inhibition immunoassay detects
IgM but not IgG anti-O9 antibodies. J Imm Meth 2003; 282: 83-91.
WHO. Background document; the diagnosis, treatment and prevention of typhoid fever.
2003; 11-16. WHO/V&B/03.07
(www.who.int/rpc/TFGuideWHO.pdf)