Citation: HUANG Zhen, XIA Shi-Qi, LIU Dao-Feng, LIU Cheng-Wei, LAI Wei-Hua. Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(2): 217-223. doi: 10.11895/j.issn.0253-3820.160572 shu

Fluorescent Microspheres Lateral Flow Assay Based on Immunomagnetic Separation for Detection of S. choleraesuis

1.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
2.
Jiangxi Academy of Forestry, Nanchang 330032, China;
3.
Jiangxi Province Centers for Disease Control and Prevention, Nanchang 330096, China

Received Date: 28 July 2016
Revised Date: 7 December 2016

Fund Project: This work was supported by the Jiangxi Agriculture Reseach System (No. JXARS-03) and the JIangxi Education Bureau Technology (No. KJLD13009)

Immunomagnetic separation (IMS) was coupled with fluorescent microspheres lateral flow assay (FM-LFA) for rapid detection of S. choleraesuis in this study. The target bacteria were firstly enriched from sample by immunomagnetic beads (IMBs), then eluted by heat treatment and detected by fluorescent microspheres lateral flow test strip. The IMBs was labeled with 30 μg/mg antibody, and the capture efficiency was greater than 90% against 10²-10⁶ CFU/mL of S. choleraesuis with great specificity. The immunofluorescent microspheres were prepared by coupling 300 μg of 11D8-D4 monoclonal antibody with 1 mg of fluorescent microspheres at pH 6. Monoclonal antibody 5F11-B11 (2.0 mg/mL) and donkey anti-mouse IgG (1.0 mg/mL) were sprayed on nitrocellulose membrane as test line and control line, respectively. The FM-LFA based on IMS was used to detect S. choleraesuis in PBS and milk. The limits of detection in PBS buffer and milk were 1.5×10⁵ CFU/mL and 7.6×10⁵ CFU/mL respectively, which were 10 and 200 times lower than that of traditional fluorescent microspheres lateral flow assay, respectively. The results showed that the method, which could enrich S. choleraesuis in milk effectively, could avoid matrix interference and improve the detection sensitivity, thus had a good application prospect.
- Immunomagnetic separation,
- Fluorescent microspheres,
- Lateral flow assay,
- S. choleraesuis
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沈阳化工大学材料科学与工程学院沈阳 110142