Abstract

A microcapillary-based loop-mediated isothermal amplification (µcLAMP) has been described for specific detection of infectious reproductive pathogens in semen samples of cattle without sophisticated instrumentation. Brucella abortus, Leptospira interrogans serovar Pomona and bovine herpesvirus 1 (BoHV-1) cultures were mixed in bovine semen samples. The µcLAMP assay is portable, user-friendly, cost-effective, and suitable to be performed as a POC diagnostic test. We have demonstrated high sensitivity and specificity of µcLAMP for detection of Brucella, Leptospira, and BoHV-1 in bovine semen samples comparable to PCR and qPCR assays. Thus, µcLAMP would be a promising field-based test for monitoring various infectious pathogens in biological samples.

• Highlights

• Detect infectious organism in bovines semen

• Reduction in carryover contamination is an important attribute, which may reduce the false-positive reaction.

• µcLAMP is a miniaturized form, which could be performed with a minimum volume of reagents.

• The µcLAMP assay is portable, user-friendly, and suitable to be performed as a POC diagnostic test.

摘要

已经描述了基于微毛细管的环介导的等温扩增（µcLAMP），无需复杂的仪器即可特异性检测牛精液样品中的传染性生殖病原体。流产布鲁氏菌，问号钩端螺旋体血清型波莫纳和牛疱疹病毒1（BoHV-1）培养物在牛精液样品中混合。µcLAMP测定法是便携式，用户友好，具有成本效益的，并且适合作为POC诊断测试进行。我们证明了µcLAMP对布鲁氏菌，钩端螺旋体，牛精液样品中的BoHV-1和BoHV-1与PCR和qPCR分析相当。因此，µcLAMP将成为一种有前途的基于现场的测试，用于监测生物样品中的各种传染性病原体。

• 强调

• 检测牛精液中的传染性生物

• 减少残留污染是一个重要的属性，可以减少假阳性反应。

• µcLAMP是一种小型化的形式，可以用最少的试剂量进行。

• µcLAMP测定法是便携式，用户友好的，适合作为POC诊断测试进行。