Standardization and Validation of Dot ELISA, Finding Detection Limit of Chlorpyrifos in Field Sprayed Samples Comparatively To Indirect Plate ELISA and Gas Chromatography
Journal Title: Scholars Journal of Applied Medical Sciences - Year 2018, Vol 6, Issue 12
Abstract
Abstract: A Dot enzyme linked immonosrbent assay (Dot ELISA) was developed and compared with standard indiect plate ELISA and gas chromatography (GC) for chlorpyrifos detection in the field samples. Firstly Dot ELISA for detection of Antichlorpyrifos antibodies (ACA) with prepared antigen and pure chlorpyrifos was standardized with the dilution range of 1µg and 1:1000µg/ml considering its limit of detection (LOD) value. Further since Dot ELISA was evaluated for field samples (fruit fodder, vegetables) for which, known dose of pure chlorpyrifos were spiked into field samples and determined with indirect plate ELISA, Dot ELISA and compared with GC. During the study four other parameters viz. sensitivity, specificity, accuracy, and K value were also determined by using Kappa statatics. Kappa statatics was used to interpret the comparative result between both types of ELISA. Results obtained from the studies showed accuracy between (50 -60%) sensitivity (10-70%) and specificity (12-90%). Percentage of accuracy above 80 considered to be positive and vice versa. While the K value i.e. % agreement between both tests lied between 0.96-0.1.0 (agreement above 0.81 showed perfect agreements). These results showed K value greater than 0.81 in all the field samples indicating perfect agreement between two techniques followed i.e. plate ELISA and Dot ELISA. This showed that both of the techniques were comparable for detection of pesticide residue detection at 75-125µg/ml dilution of antigen for Dot ELISA and 75-125ng/ml dilution of antigen among the field sprayed samples. As all the test samples showed more than 0.81 % agreement which interprets that both the test (Dot and plate ELISA) are comparable with each other. Same sample were tested with GC also which showed absence of chlorpyrifos in the field samples which supports the results obtained from the immunoassay studies. Major finding of the present studies has shown that Dot ELISA is a best tool for the qualitative detection of pesticides in the field samples with the LOD of 100µg /ml of antigen in case of chlorpyrifos.
Authors and Affiliations
Prabha Parmar
Keywords
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