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

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

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 Li  1 , Feng Chen  1
Affiliations

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

Changtao Liao et al. Front Vet Sci. .

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.

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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
Standard curves of monoplex and duplex real-time PCRs with the same targets. (A) Standard curves of monoplex and duplex real-time PCRs using the MS1 standard plasmid template. (B) Standard curves of monoplex and duplex real-time PCRs using the MS-WT standard plasmid template.
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