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基于 lipL32 和 基因的多重 PCR 分析在致病性和非致病性钩端螺旋体的分子鉴定中的应用。

Application of a Multiplex PCR Assay for Molecular Identification of Pathogenic and Non-Pathogenic Leptospires based on lipL32 and Genes.

机构信息

Department of Microbiology, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

Department of Biology, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran.

出版信息

Arch Razi Inst. 2023 Feb 28;78(1):413-418. doi: 10.22092/ARI.2022.359211.2388. eCollection 2023 Feb.

Abstract

Leptospirosis is a serious zoonotic infection and the most prevalence disease is in the tropical and subtropical region. The definitive diagnosis of Leptospirosis, caused by spirochetes of the genus infection is already using culture methods, serological tests such as the microscopic agglutination test (MAT) and molecular detection methods (PCR) are possible. In this study, we used multiplex PCR method for detection of pathogenic and non - pathogenic based on and genes. All serovars were obtained from the Reference Laboratory of Microbiology Department, Razi Vaccine and Serum Research Institute, Karaj, Iran. The PCR product for the and genes was 272 bp and 240 bp respectively. The sensitivity amplification for the multiplex assay was 10 pg / μl for gene and 10 pg / μl for gene. The sensitivity for multiplex PCR was 10 pg / μl. The results supported the idea that multiplex PCR can be used to detect samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much easily than conventional methodologies. Due to the slow growth of and the importance of time in diagnosis, molecular methods such as PCR are suggested.

摘要

钩端螺旋体病是一种严重的人畜共患病,最常见的疾病发生在热带和亚热带地区。钩端螺旋体病的明确诊断已经可以使用培养方法,也可以使用血清学检测方法(如显微镜凝集试验(MAT)和分子检测方法(PCR)。在这项研究中,我们使用基于 和 基因的多重 PCR 方法检测致病性和非致病性 。所有血清型均来自伊朗卡拉季拉齐疫苗和血清研究所微生物学系参考实验室。 和 基因的 PCR 产物分别为 272bp 和 240bp。多重检测的灵敏度扩增为 基因 10pg/μl, 基因 10pg/μl。多重 PCR 的灵敏度为 10pg/μl。结果支持了多重 PCR 可用于检测 样本的观点。该方法还能够区分腐生性和致病性钩端螺旋体,并且比传统方法更容易做到这一点。由于 的生长缓慢,以及诊断时间的重要性,建议使用 PCR 等分子方法。

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Laboratory diagnosis of leptospirosis: a challenge.钩端螺旋体病的实验室诊断:一项挑战。
J Microbiol Immunol Infect. 2013 Aug;46(4):245-52. doi: 10.1016/j.jmii.2013.03.001. Epub 2013 Apr 29.

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