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. 2024 Jun 19:15:1418959.
doi: 10.3389/fmicb.2024.1418959. eCollection 2024.

Development of a colloidal gold immunochromatographic strip for the simultaneous detection of porcine epidemic diarrhea virus and transmissible gastroenteritis virus

Jinzhu Zhou #  1   2   3 , Wei Wu #  4 , Dandan Wang #  1   2   3 , Wei Wang  1   2   3 , Xinjian Chang  1   2   3 , Yunchuan Li  1   2   3 , Jizong Li  1   2   3 , Baochao Fan  1   2   3 , Junming Zhou  1   2   3 , Rongli Guo  1   2   3 , Xuejiao Zhu  1   2   3 , Bin Li  1   2   3
Affiliations

Development of a colloidal gold immunochromatographic strip for the simultaneous detection of porcine epidemic diarrhea virus and transmissible gastroenteritis virus

Jinzhu Zhou et al. Front Microbiol. .

Abstract

In recent years, porcine diarrhea-associated viruses have caused significant economic losses globally. These viruses present similar clinical symptoms, such as watery diarrhea, dehydration, and vomiting. Co-infections with porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are common. For the rapid and on-site preliminary diagnosis on the pig farms, this study aimed to develop a colloidal gold immunochromatography assay (GICA) strip for the detection of PEDV and TGEV simultaneously. The GICA kit showed that there was no cross-reactivity with the other five common porcine viruses. With visual observation, the lower limits were approximately 104 TCID50/mL and 104 TCID50/mL for PEDV and TGEV, respectively. The GICA strip could be stored at 4°C or 25°C for 12 months without affecting its efficacy. To validate the GICA strip, 121 clinical samples were tested. The positive rates of PEDV and TGEV were 42.9 and 9.9%, respectively, and the co-infection rate of the two viruses was 5.8% based on the duplex GICA strip. Thus, the established GICA strip is a rapid, specific, and stable tool for on-site preliminary diagnosis of PEDV- and TGEV-associated diarrhea.

Keywords: RT-PCR; colloidal gold immunochromatographic strip assay; monoclonal antibodies; porcine epidemic diarrhea virus; transmissible gastroenteritis virus.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Schematic diagram of the GICA strip. (A) External structure of rapid test strip; (B) internal structure of rapid test strip; and (C) schematic diagram for reading detected results.
Figure 2
Figure 2
Identification of PEDV and TGEV monoclonal antibody. (A) The PEDV N protein was run for Western blotting using anti-PEDV McAb-1 and anti-PEDV McAb-2. (B) The TGEV N protein was run for Western blotting using anti-TGEV McAb-1 and anti-TGEV McAb-2. (C) IFA was stained for PEDV (green) and nuclei (DAPI, blue) with anti-PEDV McAb-1 and anti-PEDV McAb-2. (D) IFA was stained for PEDV (green) and nuclei (DAPI, blue) with anti-TGEV McAb-1 and anti-TGEV McAb-2.
Figure 3
Figure 3
Specificity of the GICA strip.
Figure 4
Figure 4
Minimum detection limit of the GICA strip. (A) The dilution range of the PEDV was 106–103 TCID50/mL, and the lowest detection line was 104 TCID50/mL. (B) The dilution range of the TGEV was 106–103 TCID50/mL, and the lowest detection line was 104 TCID50/mL.
Figure 5
Figure 5
Stability of the GICA strip. The kit was stable at 4 or 25°C for 1 year.
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