Standardization of an isothermal test as a novel diagnostic tool in México for the molecular detection of Chlamydia abortus in small ruminants

IF 0.5 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotecnia Pub Date : 2023-12-12 DOI:10.18633/biotecnia.v26i1.2109

Carlos Eduardo Aragon López, Erika Gabriela Palomares Resendiz, Sergio Cuevas tellechea, Javier Rolando Reyna Granados, Beatriz Arellano Reynoso, Miguel Angel Sánchez Castro, Jose Clemente Leyva Corona, Marcela Ivone Morales Pablos

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Abstract

Ovine Enzootic Abortion (OEA) is caused by Chlamydia abortus and has recently been recognized as endemic in Mexico. Due to the impact of OEA on intensive lamb production, it is necessary to establish a sanitary control plan based on an efficient diagnosis. The loop-mediated isothermal amplification technique (LAMP) is a molecular test that can specifically identify C. abortus. The aim of this study was to standardize the LAMP technique for the specific detection of C. abortus from ovine vaginal exudate. Primers were identified and modified from an electronic database. The effective conditions of the technique were determined using synthetic DNA and positive biological samples by PCR as references. The assay detection limit was 1 × 10-5 of genetic material, equivalent to 8.5 copies. The standardized LAMP test is proposed as a novel molecular tool for the rapid and specific diagnosis of C. abortus infection from ovine vaginal exudate samples in Mexico.

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在墨西哥将等温试验标准化，作为分子检测小反刍动物体内流产衣原体的新型诊断工具

绵羊流行性流产（OEA）是由流产衣原体引起的，最近被确认为墨西哥的地方病。鉴于 OEA 对集约化羔羊生产的影响，有必要在有效诊断的基础上制定卫生控制计划。环介导等温扩增技术（LAMP）是一种分子检测方法，可以特异性地鉴定流产卡氏菌。本研究的目的是将 LAMP 技术标准化，以特异性检测绵羊阴道渗出物中的流产葡萄球菌。从电子数据库中确定并修改了引物。以合成 DNA 和 PCR 阳性生物样本为参考，确定了该技术的有效条件。检测限为 1 × 10-5 的遗传物质，相当于 8.5 个拷贝。建议将标准化的 LAMP 检验作为一种新型分子工具，用于快速、特异性地诊断墨西哥绵羊阴道渗出物样本中的流产葡萄球菌感染。

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来源期刊

Biotecnia BIOTECHNOLOGY & APPLIED MICROBIOLOGY-

自引率

33.30%

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