A TaqMan-MGB real-time PCR for discriminating between MS-H-live vaccine and field Mycoplasma synoviae strains

作者：Ziqing Liu, Shouchang Zhou, Chenchen Meng, Doudou Ren, Weizhen Xiong, Guanhui Liu, Jinpeng Xu · 发表于：Microbiology Spectrum · 年份：2025 · DOI：10.1128/spectrum.03591-23 · 被引用次数：1 · 研究领域：Microbial infections and disease research、Animal Virus Infections Studies、Aquaculture disease management and microbiota

ABSTRACT Mycoplasma synoviae (MS) is an important pathogen in the poultry industry and has caused significant economic losses. Worldwide, the use of live attenuated vaccine for the MS-H strain has increased to prevent MS infection. However, there is no test available to discriminate the MS-H vaccine strains from the MS strains that are causing field infection. In this study, a TaqMan-MGB real-time PCR method (qPCR) was established, validated, and evaluated to discriminate between MS-H-live vaccine and field strains based on nucleotide differences in the hlyC gene. The validation was performed for sensitivity and reproducibility by constructing recombinant plasmids. The limits of detection were 1.07 × 10 1 copies/µL for the MS-H and 1.95 × 10 1 copies/µL for field strains, respectively. The intra- and inter-assay results were less than 2.5% based on the reproducibility test. No cross-amplification signals from other common chicken pathogens were detected. Thus, our data indicated that this qPCR is sensitive, specific, and reproducible. In addition, 709 chicken clinical samples were used to evaluate this qPCR test. The results showed that positive signals could be detected from the chicken choanal cleft swabs and are 100% in concordance with the PCR sequencing method. To the best of our knowledge, we found for the first time that both L- and C-type field MS were present in flocks immunized against the MS-H vaccine strain during the validation process. In addition, this is the f...