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Frequently asked questions for: EpiScreen™ Plus kit

FAQ1: Why was the EpiScreen kit replaced by the EpiScreen Plus kit?
The EpiScreen test measured total alpha-glucosidase, the EpiScreen Plus assay measures neutral alpha-glucosidase (NAG) activity, which is a reliable marker for epididymal function. Comparison with the WHO method has demonstrated a strong correlation between assay results.

FAQ2: Is the EpiScreen Plus method identical to the WHO method?
We have made some significant changes to the product: - (a) - The WHO method uses castanospermine to perform background correction. Castanospermine was replaced by glucose which can be stored at room temperature (versus -20°C) and which has a much greater stability - (b) - Solubility issues with one of the buffers described in the WHO manual have been resolved again resulting in a very stable and reliable product

FAQ3: Can we measure samples with a spectrophotometer, using cuvettes?
Yes, this is certainly possible but there is one main difference between both setups: One must take into account that the path length of cuvettes (most commonly 1 cm) is larger than the one of microtiter plates (usually 0.5 cm). According to the law of Lambert-Beer (Absorbance = extinction coefficient x path length x concentration), the optical density (OD) values will be approximately 2x higher. Therefore, the highest 4-nitro-phenol standards will lie outside of the measuring range of the spectrophotometer. However, based on our experience, linearity of the standard curve can be guaranteed up to 100 µM. This concentration corresponds to a neutral enzyme activity of 47.9 mIU/ml, which abundantly exceeds the cut-off value. If the activity is higher than 47.9 mIU/ml, we advise to dilute and retest the sample

FAQ4: Can we use a regular incubator for the test?
Samples must be incubated at 37°C to allow optimal enzyme activity. Therefore, we advise to use a thermoshaker or a warm water bath, where samples are in direct contact with the heated environment. A standard cell culture incubator is not suited for this purpose, as heat transfer through the air is slower, hereby resulting in suboptimal reaction conditions.

FAQ5: Can results of the EpiScreen Plus test be compared one on one with those with the EpiScreen test?
No. The old EpiScreen assay measured total alpha-glucosidase activity, which consists of a neutral (epididymis-related) and acid (prostate-derived) fraction.
The new kit is designed to assess neutral activity in very specific and reliable manner. Assay outcome will therefore be lower than or equal to the total activity.
The old assay was designed as a screening tool to distinguish samples with normal and abnormal epididymis function.

FAQ6: Do we need to prepare a standard curve with each test?
Yes

FAQ7: How do we calculate the alpha-glucosidase value?
We have developed and validated an MS Excel™ template for automatic data processing.
The template is available upon request, please send an e-mail to epitemplate@fertipro.com

Resources

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