Citation: REN Shuai, DING Qiao-Qi, LI Li, SHAO Man-Yu, SONG Su-Quan, YAN Li-Ping. Development and Application of A Novel Chemiluminescence Chip Immunoassay for Detection of Chloramphenicol[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(10): 1581-1587. doi: 10.11895/j.issn.0253-3820.181189 shu

Development and Application of A Novel Chemiluminescence Chip Immunoassay for Detection of Chloramphenicol

REN Shuai ^1,2,3, ,
DING Qiao-Qi ^1,2 ,
LI Li ^1,2 ,
SHAO Man-Yu ^1,2 ,
SONG Su-Quan ¹ ,
YAN Li-Ping ^1,2,3,4

1.
College of Veterinary Medicine;
2.
MOE Joint International Research Laboratory of Animal Health and Food Safety;
3.
Jiangsu Engineering Laboratory of Animal Immunology, Institute of Immunology;
4.
Jiangsu Detection Center of Terrestrial Wildlife Disease, Institute of Immunology, Nanjing Agricultural University, Nanjing 210095, China

Received Date: 23 March 2018
Revised Date: 30 August 2018

Fund Project: This work was supported by the National Key R&D Program (No. 2016YFD0501200), the Fundamental Research Funds for the Central Universities (No. KYZ201848), the Natural Science Foundation of Jiangsu Province (No. BK20161452), and the Key Program of Science and Technology Planning of Guangdong Province, China (No. 2017B020202010).

The rabbit anti-chloramphenicol (CAP) polyclonal antibodies were prepared by immunizing New Zealand rabbits by CAP antigen (CAP-HS-BSA), with antibody titer of 2×10⁵. Then the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and chemiluminescence immunoassay based on iPDMS for detection of CAP were established. The proposed ic-ELISA showed a half-inhibition concentration (IC₅₀) of 12.9 ng/mL, with a linear detection range (IC₂₀-IC₈₀) of 0.8-250.0 ng/mL and limit of detection (LOD, IC₁₀) of 0.1 ng/mL. The standard curve was B/B₀=-0.2028lgC_CAP+0.7281(R²=0.9926). The cross-reactivity of ic-ELISA towards structural analogues of CAP was less than 0.01%. When applied to detection of CAP in real milk sample, the average recoveries of ic-ELISA were from 81.8% to 97.9%, with relative standard deviation (RSD) of less than 10%. However, for the proposed chemiluminescence immunoassay, the IC₅₀ was 263.0 ng/mL, the linear range (IC₂₀-IC₈₀) was 1-5000 ng/mL, and the LOD was 1 ng/mL. The standard curve was B/B₀=-0.1781lgC_CAP +0.9318(R²=0.9891). The average recoveries of iPDMS based chemiluminescence immunoassay were from 77.0% to 100.5%, with the RSD of less than 15% for spiked milk samples.
- Chloramphenicol,
- Polyclonal antibody,
- Indirect competitive enzyme-linked immunosorbent assay,
- Chemiluminescence,
- Chip immunoassay
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