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. 2017 Apr 29;9(5):152.
doi: 10.3390/toxins9050152.

Development of an Immunochromatographic Strip Test for the Rapid Detection of Alternariol Monomethyl Ether in Fruit

Yan Man  1   2   3 , Gang Liang  4   5   6 , Fuchao Jia  7 , An Li  8   9   10 , Hailong Fu  11   12   13 , Meng Wang  14   15   16 , Ligang Pan  17   18   19
Affiliations

Development of an Immunochromatographic Strip Test for the Rapid Detection of Alternariol Monomethyl Ether in Fruit

Yan Man et al. Toxins (Basel). .

Abstract

A rapid, portable, and semi-quantitative immunochromatographic strip was developed for rapid and visual detection of alternariol monomethyl ether (AME). For this purpose, the anti-AME monoclonal antibody (mAb) was prepared and identified. AME coupled to bovine serum albumin (BSA) via methyl 4-bromobutanoate was prepared as immunogen. The recoveries of AME in spiked cherry and orange fruits determined by competitive ELISA were 86.1% and 80.7%, respectively. A colloidal gold nanoparticle (CGN) and CGNs-mAb conjugate were synthesized, and on this basis, a competitive colloidal gold immunochromatographic strip was developed and applied to the detection of AME toxin in fruit samples. The intensity of red density of the test line (T line) is inversely proportional to AME concentration in the range 0.1-10 ng/mL. The visual limit of detection (LOD) of AME was found to be about 10 ng/mL. The semi-quantitative detection can be completed in 10 min. Moreover, the immunochromatographic strip has lower cross-reactivity with AME analogues, and it has a good stability performance (following 3 months of storage). Hence, the colloidal gold immunochromatographic strip could be used as a semi-quantitative tool for the on-site, rapid, and visual detection of AME in fruit.

Keywords: alternariol monomethyl ether (AME); colloidal gold nanoparticle; competitive ELISA; immunochromatographic strip test; monoclonal antibody.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The schematic preparation of carboxyl derivative modification of alternariol monomethyl ether (AME).
Figure 2
Figure 2
Typical standard curves of the competitive ELISA for alternariol monomethyl ether (AME). Three replicate wells of all the standard concentrations (0.01, 0.1, 0.3, 0.9, 2.7, 8.1 ng/mL) were analyzed.
Figure 3
Figure 3
TEM image of colloidal gold nanoparticles (CGNs) (a) and UV spectrum of the CGNs-mAb conjugates (b). The black line and red line in b represent CGNs and CGNs-mAb conjugates, respectively.
Figure 4
Figure 4
Schematic illustration of immunochromatographic strip. The T line (test line) and C line (control line) was coated by AME- ovalbumin (OVA) and goat anti-mouse IgG, respectively.
Figure 5
Figure 5
Limit of detection (LOD) of AME with colloidal gold immunochromatographic strip. A series of dilutions (0–50 ng/mL) of AME standard solutions were prepared by diluting AME in PBS. When the concentration of AME was higher than 10 ng/mL, the red line at the T line disappeared.
Figure 6
Figure 6
Cross reactivity of the immunochromatographic strip with alternariol (AOH), altenuene (ALT), and tenuazonic acid (TeA).
Figure 7
Figure 7
The AME test results of immunochromatographic strips after storage for 3 months at room temperature. The concentrations of AME were 0, 0.1, 1, 5, 10, 20, 30, and 40 ng/mL, respectively.
Figure 8
Figure 8
Immunochromatographic strip test of AME in spiked cherry and orange samples. The spiked concentrations of AME in cherry were 0, 1, 5, 10, and 20 ng/mL, and in orange were 0, 1, and 10 ng/mL.
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