Abstract：
Mastitis is a common mammary gland disease of dairy cattle caused by a wide range of organisms including bacteria, fungi and algae. Mastitis contributes to economic losses of dairy farms due to reduced yield and poor quality of milk. Since the correct identification of pathogens responsible for the development of mastitis is crucial to the success of treatment, it is necessary to develop a quick and accurate test to distinguish the main pathogens causing this disease. In this paper, we describe the development of a test based on the multiplex polymerase chain reaction (PCR) method allowing for the identification of Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Staphylococcus aureus. When creating our test, we relied on the results from new generation sequencing (NGS) for accurate determination of species affiliation. The multiplex PCR test was verified on 100 strains including veterinary samples, ATCC and Polish Collection of Microorganisms (PCM) reference strains. The obtained results indicate that this test is accurate and displays high specificity. It may serve as a valuable molecular tool for the detection of major mastitis pathogens.
摘要：
乳腺炎是奶牛常见的乳腺疾病，由包括细菌在内的多种生物引起，真菌和藻类。由于产量降低和牛奶质量差，乳腺炎导致奶牛场的经济损失。由于正确鉴定导致乳腺炎发展的病原体对治疗的成功至关重要，有必要开发一种快速准确的测试方法来区分引起这种疾病的主要病原体。在本文中,我们描述了基于多重聚合酶链反应（PCR）方法的测试的发展，允许鉴定无乳链球菌，乳酸链球菌，乳房链球菌和金黄色葡萄球菌。在创建我们的测试时，我们依靠新一代测序(NGS)的结果来准确确定物种隶属度.多重PCR检测在包括兽医样本在内的100个菌株上进行了验证，ATCC和波兰收集微生物(PCM)参考菌株。获得的结果表明该测试是准确的并且显示出高特异性。它可以作为检测主要乳腺炎病原体的有价值的分子工具。
