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Ct value-based real time PCR serotyping of Glaesserella parasuis

文献类型： 外文期刊

作者： Cui, Yifang 1 ; Guo, Fangfang 1 ; Cai, Xuwang 2 ; Cao, Xiaoya 1 ; Guo, Jie 1 ; Wang, Hongjun 1 ; Yang, Bing 1 ; Zhou, Hon 1 ;

作者机构： 1.Beijing Acad Agr & Forestry Sci, Inst Anim Husb & Vet Med, Beijing Key Lab Prevent & Control Infect Dis Live, Beijing 100097, Peoples R China

2.Huazhong Agr Univ, Div Anim Infect Dis, State Key Lab Agr Microbiol, Wuhan 430070, Hubei, Peoples R China

3.Univ Queensland, Queensland Alliance Agr & Food Innovat, St Lucia, Qld 4072, Australia

关键词： Glaesserella parasuis; Molecular serotyping; Real time PCR; Cycle threshold

期刊名称：VETERINARY MICROBIOLOGY （ 影响因子：3.03； 五年影响因子：2.923 ）

ISSN： 0378-1135

年卷期： 2021 年 254 卷

页码：

收录情况： SCI

摘要： Glaesserella parasuis is the causative agent of Glasser's disease in swine. Serotyping plays an essential role in prevalence investigations and in the development of vaccination strategies for the prevention of this disease. Molecular serotyping based on variation within the capsule loci of the 15 serovars is more accurate and efficient than traditional serological serotyping. To reduce the running time and facilitate ease of data interpretation, we developed a simple and rapid cycle threshold (Ct) value-based real time PCR (qPCR) method for the identification and serotyping of G. parasuis. The qPCR method distinguished between all 15 serovar reference strains of G. parasuis with efficiency values ranging between 85.5 % and 110.4 % and, R-2 values > 0.98. The qPCR serotyping was evaluated using 83 clinical isolates with 43 of the isolates having been previously assigned to a serovar by the gel immuno-diffusion (GID) assay and 40 non-typeable isolates. The qPCR results of 41/43 (95.3 %) isolates were concordant with the GID assay except two isolates of serovar 12 were assigned to serovar 5. In addition, the qPCR serotyping assigned a serovar to each of the 40 non-typeable isolates. Of the 83 isolates tested to assign a serovar, a concordance rate of 98.8 % (82/83) was determined between the qPCR and the previously reported multiplex PCR of Howell et al. (2015) (including those that were either serovars 5 or 12). Despite the inability to differentiate between serovars 5 and 12, the Ct value-based qPCR serotyping represents an attractive alternative to current molecular serotyping method for G. parasuis and could be used for both epidemiological monitoring and the guidance of vaccination programs.

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