Dissertation > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary Pharmacology > Veterinary Toxicology > Chemical residues in animal foods

Dot-immunogold Filtration Assay for Rapid Detection of Sulfamethoxazole Residues

Author JiangHong
Tutor WuGuoJuan;HuangYan
School Xinjiang Agricultural University
Course Basic Veterinary Science
Keywords Sulphamethoxazole Polyclonal antibody dot-immunogold filtration assay
CLC S859.84
Type Master's thesis
Year 2004
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Polyclonal antibody(PcAb) of Sulphamethoxazole(SMZ),Sulfanilamide(SN),Sulfadimethoxine(SDM) were prepared, The establishment of dot-immunogold filtration assay(DIGFA) for rapid dection of sulfamethoxazole residues would be a base for the use of DIGFA in dectecting residues of sulfanomides.Hapten SMZ,SN,SDM were conjugated to human serum albumin (HSA),ovalbumin(OVA)or rabbit serum albumin(RSA) by diazotization for preparing complete antigen SMZ-HSA, SMZ-RSA, SN-HSA, SDM-HSA, SMZ-OVA, SN-OVA, SDM-OVA. The results showed a successful coupling occurrence with ultraviolet(UV) scanning spectrum and high performance liquid chromatography(HPLC). Rabbits had been immunized respectively with SMZ-HSA, SMZ-RSA, SN-HSA, SDM-HSA, and mouse had been immunized with SMZ-HSA. The results showed that high titer and specificity antiserums from those rabbits and mouse were testified by double agar diffusion and indirect ELISA.And they had no cross-reactivity with OVA by indirect ELISA.The results of competitive indirect ELISA showed that the sensitivity to correspond sulfonomides ofPcAbs exhibited from those rabbits were 46.618ng/ml,22.442ng/ml,14.0086ng/ml,73.9970ng/ml, respectively.PcAb showed high affinity and specificity for correspond sulfonomides and low affinity for other seven structurally related sulfonomides.Results of purified SMZ-HSA rabbit PcAb Sodium dodecyl sulfate- Polyacrylamide gel electrophoresis (SDS –PAGE) suggested that purified PcAb was electrophoretic pure.DIGFA for rapid dection of sulfamethoxazole residues was established by using two match PcAb prepared in our laboratory.one PcAb was inoculated on nitrocellulose membrane,and another was conjugated with colloidal gold. The sensitivity of this method could reach 2.5ng/dot and no cross reaction with other seven structurally related sulfonomides.The indentical rate to HPLC was 93.01%.

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