Development of a rapid quantitative method to differentiate MS1 vaccine strain from wild-type Mycoplasma synoviae

Changtao Liao; Yiquan Chen; Zhuanqiang Yan; Yiwei Song; Qi Zhou; Puduo Zhu; Xudong He; Wenyang Li; Feng Chen

doi:10.3389/fvets.2024.1354548

Development of a rapid quantitative method to differentiate MS1 vaccine strain from wild-type Mycoplasma synoviae

Front Vet Sci. 2024 Mar 1:11:1354548. doi: 10.3389/fvets.2024.1354548. eCollection 2024.

Authors

Changtao Liao^#^{1

2}, Yiquan Chen^#¹, Zhuanqiang Yan^{1

2}, Yiwei Song^{1

2}, Qi Zhou^{1

2}, Puduo Zhu^{1

2}, Xudong He^{1

2}, Wenyang Li¹, Feng Chen¹

Affiliations

¹ College of Animal Science, South China Agricultural University, Guangzhou, China.
² Yunfu Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Yunfu, China.

^# Contributed equally.

Abstract

Mycoplasma synoviae (MS) is an economically important pathogen in the poultry industry. Vaccination is an effective method to prevent and control MS infections. Currently two live attenuated MS vaccines are commercially available, the temperature-sensitive MS-H vaccine strain and the NAD-independent MS1 vaccine strain. Differentiation of vaccine strains from wild-type (WT) strains is crucial for monitoring MS infection, especially after vaccination. In this study, we developed a Taqman duplex real-time polymerase chain reaction (PCR) method to identify MS1 vaccine strains from WT strains. The method was specific and did not cross-react with other avian pathogens. The sensitivity assay indicated that no inhibition occurred between probes or between mixed and pure templates in duplex real-time PCR. Compared with the melt-based mismatch amplification mutation assay (MAMA), our method was more sensitive and rapid. In conclusion, the Taqman duplex real-time PCR method is a useful method for the diagnosis and differentiation of WT-MS and MS1 vaccine strains in a single reaction.

Keywords: MS attenuated live vaccines; Mycoplasma synoviae; differentiation methods; real-time PCR; wild-type strains.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the China Postdoctoral Science Foundation [grant number 2021M692455].