RIDA TUBE Calprotectin
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1 RIDA TUBE Calprotectin Accessory for RIDASCREEN Calprotectin Art. No. GZ3016 R-Biopharm AG, An der neuen Bergstraße 17, D Darmstadt, Germany Tel.: +49 (0) / Telefax: +49 (0)
2 1. Intended use For in-vitro diagnostics. RIDA TUBE Calprotectin is used for the sampling and the homogenization of human stool samples in the laboratory. They are accessories for the RIDASCREEN Calprotectin ELISA assay G Kit content One kit contains 50 stool sampling tubes. 3. Description of stool sampling tube One RIDA TUBE is composed of the following parts: - tube - measuring stick with measuring tip - Funnel 4. Storage The stool collection tubes need to be stored at 2 8 C. 5. Necessary equipment Vortex, inoculation loop or wooden stick 2
3 6. Warning/ precautions We do not recommend centrifuging the tube. Stool samples should be treated as potentially infectious material. 7. Collection and storage of samples Stool samples should be shipped cooled if possible and stored at 2 8 C. If the analysis will not start immediately after arrival of the samples (within 3 days), storage at -20 C or colder is recommended. Repeated freezing and thawing of samples should be avoided. 7.1 Liquid stool 10 µl of liquid stool samples could be pipetted directly into the stool collection tube. 7.2 Very solid stool Very solid stool should be thoroughly homogenized before transfer into the stool collection tube, e.g. by using a wooden stick or inoculation loop. In case the stool is very hard, besides vortexing the tube it is recommended to tip the tube softly onto a hard surface until the sample is fully removed from the measuring tip. 8. Sample preparations The stool sample should be at room temperature (20 25 C) before extraction and should be homogenized e.g. with an inoculation loop or a wooden stick. When transferred to the stool collection tube it should be taken care, that the grooves of the measuring tip are completely filled with stool. Moreover, no stool should be at the bar of the measuring stick. 3
4 9. Collecting samples with sample collection device instruction for use 1. Unscrew the measuring stick (orange cap). 2. Remove the stick with the measuring tip. 3. Dip the measuring tip into the stool specimen at three different places. 4. Make sure that the grooves of the measuring tip are filled with stool. 5. Return the stick into the tube. By inserting the stick into the tube, excess stool material remains in the blue funnel insert. Close the tube carefully. The measuring stick collects approx. 10 mg of stool sample. 10 μl of liquid stool samples could be pipetted into the stool collection tube. 6. Shake the tube by vortexing prior to testing. The stool material has to be suspended completely in extraction buffer. In case the stool is very hard it is recommended to tip the tube softly onto a hard surface until the sample is fully removed from the measuring tip. 7. Let the stool extract sediment. The RIDA TUBE Calprotectin shouldn t be centrifuged. The stool extract cannot be stored and should be used for analysis directly after dilution. 8. For testing, open the tube at the blue shutter. Dilute 100 μl of the stool suspension in 900 μl RIDASCREEN Sample dilution buffer (Diluent 3). 100 μl of the (1:10) diluted stool sample can be directly used in the assay. Note: The RIDA TUBE Calprotectin can also be used on automated 4-plate ELISA- Readers, e.g. Dynex DSX. Sedimentation time of 15 minutes can help to avoid possible difficulties with dispension. 10. Storage of extracts The calprotectin concentrations in the extraction buffer of the RIDA TUBE Calprotectin (GZ3016) are only partially stable for 7 days at 4 C. For this reason it is recommended to use the fresh extract immediately in the test. However the majority of analyzed stool extracts of the RIDA TUBE Calprotectin (GZ3016) showed a stabile calprotectin concentration for a period of at least 3 days. Only 3 out of 78 samples (3.8 %) showed a significant increase in calprotectin concentration. 1 out of 78 samples (1.3 %) showed a significant decrease in calprotectin concentration. None of the analyzed samples, which showed a calprotectin concentration well below the cut off of 50 mg/kg and were tested as negative, became distinctly positive after storage of 7 days. 4
5 11. Precision data 11.1 Comparison of the sample extraction using a sample tube or weighing 39 stool samples in a concentration range of to mg/kg were tested with RIDASCREEN Calprotectin. The stool volume was collected either, as described in the IFU by weighing or through sampling by RIDASCREEN Stuhlröhrchen. The concentrations were determined for both methods and the correlation coefficient (r 2 ) of r² = 0.99 was calculated (see figure 1). Figure 1: Correlation of Calprotectin concentrations after sampling by RIDASCREEN Stuhlröhrchen or by weighing. The correlation coefficient is r² = The correlation coefficient of r²= 0.99 shows the high comparability of the results of weighing versus RIDASCREEN Stuhlröhrchen Equivalence between RIDASCREEN Stuhlröhrchen (GZ3006) and RIDA TUBE Calprotectin (GZ3016) 40 stool samples in a concentration range of to mg/kg were tested with RIDASCREEN Calprotectin. The stool volume was collected either, as described in the IFU taken with RIDA TUBE Calprotectin (GZ3016) or RIDASCREEN Stuhlröhrchen (GZ3006). The concentrations were determined for both methods and the correlation coefficient (r2) of r² = 0.96 was calculated (see figure 1). 5
6 The correlation coefficient of r²= 0.96 shows the high comparability of the results of RIDASCREEN Stuhlröhrchen (GZ3006) versus RIDA TUBE Calprotectin (GZ3016). The data regarding the equivalence clearly show the high correlation of the sample taking between weighing and RIDASCREEN Stuhlröhrchen, as well as between RIDASCREEN Stuhlröhrchen and RIDA TUBE Calprotectin. This demonstrates that RIDA TUBE Calprotectin is also suited for the sample taking with RIDASCREEN Calprotectin. 6
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