The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of natamycin in biological agents.
1. For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.
It is highly recommended to use the remaining reagents within 1 month provided this is prior to the expiration date of the kit. For the expiration date of the kit, please refer to the label on the kit box. All components are stable up to the expiration date.
Biological agents - Remove particulates by centrifugation and assay immediately or aliquot and store samples at ≤-20°C. Avoid repeated freeze-thaw cycles.
1. Samples to be used within 5 days may be stored at 4°C, otherwise samples must be stored at -20°C (≤1 month) or -80°C (≤2 months) to avoid loss of bioactivity and contamination.
2. When performing the assay, bring samples to room temperature.
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level natamycin were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level natamycin were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
3.70-300ng/mL. The standard curve concentrations used for the ELISA's were 300ng/mL, 100ng/mL, 33.33ng/mL, 11.11ng/mL, 3.70ng/mL.
The minimum detectable dose of natamycin is typically less than 1.45ng/mL.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by subtracting two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.
Natamycin, also known as pimaricin and sometimes sold as Natacyn, is a naturally occurring antifungal agent produced during fermentation by the bacterium Streptomyces natalensis, commonly found in soil. Natamycin has a very low solubility in water; however, natamycin is effective at very low levels. Its minimum inhibitory concentration is less than 10 ppm for most molds. Natamycin is classified as a macrolide polyene antifungal, and as a drug, is used to treat fungal keratitis, an infection of the eye. It is especially effective against Aspergillus and Fusarium corneal infections. Other common members of the polyene macrolide antifungal family are amphotericin B, nystatin, and filipin. Natamycin is also used in the food industry as a natural preservative.