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2012-2019 年德国分离的格氏李斯特菌的血清分型和病原分型比较不同基于 PCR 的方法。

Serotyping and pathotyping of Glaesserella parasuis isolated 2012-2019 in Germany comparing different PCR-based methods.

机构信息

Institute for Microbiology, Department of Infectious Diseases, University of Veterinary Medicine, Foundation, Hannover, Germany.

Clinic for Swine and Small Ruminants and Forensic Medicine and Ambulatory Service, University of Veterinary Medicine Hannover, Foundation, Hannover, Germany.

出版信息

Vet Res. 2020 Nov 17;51(1):137. doi: 10.1186/s13567-020-00862-1.

Abstract

Glaesserella parasuis is an important pathogen in swine production. It acts as a primary pathogen in systemic Glässer´s disease and as a secondary pathogen in Porcine Respiratory Disease Complex. In this study, a collection of 308 isolates from carrier animals and individuals with respiratory or Glässer´s disease isolated 2012-2019 in Germany was analysed. Isolates were characterized for serovar implementing two different PCR methods. Additionally, two different PCR methods for pathotyping isolates were applied to the collection and results compared. Serovar 6 (p < 0.0001) and 9 (p = 0.0007) were correlated with carrier isolates and serovar 4 was associated with isolates from animals with respiratory disease (p = 0.015). In systemic isolates, serovar 13 was most frequently detected (18.9%). Various other serovars were isolated from all sites and the ratio of serovar 5 to serovar 12 was approximately 1:2. These two serovars together represented 14.3% of the isolates; only serovar 4 was isolated more frequently (24.7%). The pathotyping method based on the leader sequence (LS = ESPR of vta) was easy to perform and corresponded well to the clinical background information. Of the carrier isolates 72% were identified as non-virulent while 91% of the systemic isolates were classified as virulent (p < 0.0001). Results of the pathotyping PCR based on 10 different marker genes overall were in good agreement with clinical metadata as well as with results of the LS-PCR. However, the pathotyping PCR was more complicated to perform and analyze. In conclusion, a combination of the serotyping multiplex-PCR and the LS-PCR could improve identification of clinically relevant G. parasuis isolates, especially from respiratory samples.

摘要

副猪嗜血杆菌是猪生产中的一种重要病原体。它既是全身性 Glässer 病的主要病原体,也是猪呼吸道疾病复合症的次要病原体。在本研究中,对 2012 年至 2019 年期间从德国带菌动物和患有呼吸道或 Glässer 病的个体中收集的 308 株分离株进行了分析。使用两种不同的 PCR 方法对分离株进行血清型分析。此外,还应用两种不同的 PCR 方法对分离株进行病原分型,并对结果进行比较。血清型 6(p<0.0001)和 9(p=0.0007)与带菌分离株相关,血清型 4 与患有呼吸道疾病的动物分离株相关(p=0.015)。在系统性分离株中,血清型 13 最为常见(18.9%)。从所有部位还分离到各种其他血清型,血清型 5 与血清型 12 的比例约为 1:2。这两种血清型共占分离株的 14.3%;仅血清型 4 的分离频率更高(24.7%)。基于 leader 序列(LS=ESPR of vta)的病原分型方法易于操作,与临床背景信息吻合良好。在带菌分离株中,72%被鉴定为非毒力株,而在系统性分离株中,91%被鉴定为毒力株(p<0.0001)。基于 10 个不同标记基因的病原分型 PCR 结果与临床元数据以及 LS-PCR 结果总体上一致。然而,病原分型 PCR 操作和分析更为复杂。总之,血清型多重 PCR 和 LS-PCR 的结合可以提高对临床相关副猪嗜血杆菌分离株的鉴定,尤其是来自呼吸道样本的鉴定。

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