sažetak izlaganja sa skupa
Recently published method validation reported increased selectivity of tetramethyl-benzidine (TMB)-based presumptive tests, by adding 0.5M ethylendiaminetetraacetic acid (EDTA) to a reaction pad. Chelator EDTA binds positively charged metal ions and other substances preventing them to react with TMB in pad, i. e. eliminating possible false positives. Hence, the main goal of this study was to verify these data for subsequent upgrade of ours validated protocols. In addition, possible interference of the 0.5M EDTA on DNA analysis was evaluated.
Twenty one substances/surfaces were tested. Whole blood (known donor) was used as a positive control, and test strips moistened with ultrapure water as negative control. All testing were done in triplicates, with methods of EDTA application previously described. Presence/absence of colored reaction was recorded at 30s and 60s and interpreted as positive or negative after 60s.
Rust, brass, brick, soil-1, tomato pureè/paste and saliva samples of known donor exhibited negative results for test strips moistened with water and 0.5M EDTA. Contrary to previously published, majority of tested samples including plant leaf, copper surface and bleach, showed false positives with 0.5M EDTA. Negative results with EDTA for wood branch, soil-2 and banana pointed out possible influence of EDTA on increased selectivity, since positive results were obtained with water. Finally, all EDTA tested blood samples resulted in complete DNA profile using the GlobalFiler™ PCR Amplification Kit.
Overall findings showed considerable discrepancy when compared with previously published, indicating that further study about influence of EDTA on selectivity of Combur3 Test® E are required. Sodium hypochlorite as a major compound of comercially available bleaches, still remains important substance causing false positives. Nevertheless, Combur3 Test® E is distinguished as highly sensitive, cheap, easy to use and STRs analysis compatible presumptive test for blood.
blood; Combur3 Test® E; selectivity; EDTA; STRs analysis
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