Molecular Detection of Some Most Important Pathogenic Bacteria by Multiplex PCR for Diagnosis of Subclinical Mastitis in Bovine

Sudeep Solanki, Durga Devi
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Abstract

Background: The dairy sector faces formidable obstacles in the early detection of the bacteria causing subclinical mastitis and in the proactive treatment of those cases. The objective of the study was to access multiplex PCR assays (MPCR) and conventional methods for the concurrent identification of significant bacteria that cause sub-clinical mastitis and to compare these methods. 200 samples of pooled milk from cows and buffaloes in Rajasthan were gathered between 2020 and 2021. Methods: The primary bacterial pathogens in milk samples were identified using conventional methods such multiplex PCR, biochemical testing and culture. Result: From 200 combined samples of cow and buffalo milk, the traditional approach was able to extract 97 different strains. Staphylococcus aureus 54 (27%), Streptococcus spp. 30 (15%) and E. coli 13 (6.5%) were shown to be prevalent as single or mixed infections, respectively. Staphylococcus was the main pathogen that was discovered. concurrently, S. aureus, Streptococcus and E. coli. Direct detection of Staphylococcus 65 (32.5%), Streptococcus 37 (18.5%), E. coli and 16 (8%) by multiplex PCR was found in milk samples. The analysis revealed that because multiplex PCR assays have higher specificity and sensitivity than conventional procedures, they are more reliable. The multiplex PCR method employed in the current study was a simple and quick technique to identify the major pathogens and it has the potential to be a very helpful tool for determining the pathogens that cause environmental mastitis and evaluating the health of the herd.
利用多重 PCR 对一些最重要的致病菌进行分子检测,以诊断牛的亚临床乳腺炎
背景:乳制品行业在早期检测引起亚临床乳腺炎的细菌和积极治疗这些病例方面面临着巨大障碍。本研究的目的是采用多重 PCR 检测法(MPCR)和传统方法同时鉴定引起亚临床乳腺炎的重要细菌,并对这些方法进行比较。在 2020 年至 2021 年期间收集了 200 份拉贾斯坦邦奶牛和水牛的牛奶样本。方法:使用多重 PCR、生化测试和培养等传统方法鉴定牛奶样本中的主要细菌病原体。结果从 200 份牛奶和水牛奶的组合样本中,传统方法能够提取出 97 种不同的菌株。结果表明,金黄色葡萄球菌 54 株(27%)、链球菌 30 株(15%)和大肠杆菌 13 株(6.5%)分别为单一或混合感染。金黄色葡萄球菌、链球菌和大肠杆菌同时也是发现的主要病原体。通过多重 PCR 技术在牛奶样本中直接检测出葡萄球菌 65 个(32.5%)、链球菌 37 个(18.5%)、大肠杆菌 16 个(8%)。分析表明,与传统方法相比,多重 PCR 方法具有更高的特异性和灵敏度,因此更加可靠。本研究中采用的多重 PCR 方法是一种简单快速的主要病原体鉴定技术,有可能成为确定引起环境性乳腺炎的病原体和评估牛群健康状况的一种非常有用的工具。
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