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Using a novel gene site to develop a duplex real-time TaqMan MGB probe PCR method for the SNP detection and differentiation between the MS-H live vaccine strain and wild-type Mycoplasma synoviae strains

文献类型： 外文期刊

作者： Zhao, Luru 1 ; Tang, Xiaochuan 2 ; Guo, Weiqi 1 ; Zhang, Bin 1 ; Peng, Haoheng 1 ; Ye, Lijun 1 ; Liu, Yinan 1 ; Liang, Jingyi 1 ; Tian, Mingxing 1 ; Bao, Yanqing 1 ; Qi, Jingjing 1 ; Wang, Shaohui 1 ;

作者机构： 1.Chinese Acad Agr Sci CAAS, Shanghai Vet Res Inst, 518 Ziyue Rd, Shanghai 200241, Peoples R China

2.Guangxi Univ, Coll Anim Sci & Technol, 100 East Univ Rd, Nanning 530004, Peoples R China

关键词： Mycoplasma synoviae; Quantitative PCR; ktrB; SNP; MGB

期刊名称：POULTRY SCIENCE （ 影响因子：4.2； 五年影响因子：4.5 ）

ISSN： 0032-5791

年卷期： 2025 年 104 卷 5 期

页码：

收录情况： SCI

摘要： Mycoplasma synoviae (MS) is a globally prevalent avian pathogen responsible for airsacculitis and synovitis. The temperature-sensitive (ts)+ vaccine strain MS-H, a live attenuated variant, is the most effective and widely used vaccine for controlling infections in the poultry industry. Consequently, accurate detection is essential for a strategy known as differentiating infected from vaccinated animals (DIVA). In this study, we developed a duplex real-time TaqMan minor groove binder (MGB) probe PCR (The DRTM-probe PCR) method to differentiate the MS-H live vaccine strain from wild-type strains by targeting a single nucleotide polymorphism (SNP) in the ktrB gene. This gene overcomes the restoration of the genotype of wild-type 86079/7NS in specific regions. With a detection limit of 6.25 copies/mu L, the DRTM-probes PCR method demonstrates a good specificity in distinguishing in one hour. For simulated clinical samples, the method achieved over 95 % sequence identity with reference fragments, confirming its accuracy. The established DRTM-probe PCR method offers a specific, rapid, and reliable approach for SNP detection with significant application potential.

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