Abstract:
Human brucellosis caused by Brucella is a widespread zoonosis that is prevalent in many countries globally. The high homology between members of the Brucella genus and Ochrobactrum spp. often complicates the determination of disease etiology in patients. The efficient and reliable identification and distinction of Brucella are of primary interest for both medical surveillance and outbreak purposes. A large amount of genomic data for the Brucella genus was analyzed to uncover novel probes containing single-nucleotide polymorphisms (SNPs). GAMOSCE v1.0 software was developed based on the above novel eProbes. In conjunction with clinical requirements, an RPA-Cas12a detection method was developed for the on-site determination of B. abortus and B. melitensis by fluorescence and lateral flow dipsticks (LFDs). We demonstrated the potential of these probes for rapid and accurate detection of the Brucella genus and five significant Brucella species in silico using GAMOSCE. GAMOSCE was validated on different Brucella datasets and correctly identified all Brucella strains, demonstrating a strong discrimination ability. The RPA-Cas12a detection method showed good performance in detection in clinical blood samples and veterinary isolates. We provide both in silico and on-site methods that are convenient and reliable for use in local hospitals and public health programs for the detection of brucellosis.
摘要:
由布鲁氏菌引起的人类布鲁氏菌病是一种广泛的人畜共患病,在全球许多国家都很普遍。布鲁氏菌属和苍白杆菌属之间的同源性很高。通常会使患者疾病病因的确定复杂化。布鲁氏菌的有效和可靠的识别和区分对于医学监测和爆发目的都具有重要意义。分析了布鲁氏菌属的大量基因组数据,以发现含有单核苷酸多态性(SNP)的新型探针。GAMOSCEv1.0软件是基于上述新颖的eProbes开发的。结合临床要求,开发了一种RPA-Cas12a检测方法,用于通过荧光和侧流试纸(LFDs)现场测定B.abortus和B.melitensis。我们证明了这些探针在使用GAMOSCE快速准确检测布鲁氏菌属和五种重要的布鲁氏菌属的潜力。GAMOSCE在不同的布鲁氏菌数据集上进行了验证,并正确识别了所有布鲁氏菌菌株,具有很强的鉴别能力。RPA-Cas12a检测方法在临床血液样品和兽医分离株中显示出良好的检测性能。我们提供计算机和现场方法,方便可靠,适用于当地医院和公共卫生计划,以检测布鲁氏菌病。