通过靶向和基因的封闭环介导等温扩增法在人类临床样本中快速可视化检测耐甲氧西林菌。
Rapid Visual Detection of Methicillin-Resistant in Human Clinical Samples via Closed LAMP Assay Targeting and Genes.
作者信息
Abusheraida Noora S A, AlBaker Asraa A H, Aljabri Asmaa S A, Abdelrahman Hana A, Al-Mana Hassan, Wilson Godwin J, Anan Khalid A, Eltai Nahla O
机构信息
College of Health Science, Qatar University, Doha P.O. Box 2713, Qatar.
Biomedical Research Center, Qatar University, Doha P.O. Box 2713, Qatar.
出版信息
Microorganisms. 2024 Jan 12;12(1):157. doi: 10.3390/microorganisms12010157.
The emergence of antimicrobial resistance (AMR), particularly methicillin-resistant (MRSA), poses a significant global health threat as these bacteria increasingly become resistant to the most available therapeutic options. Thus, developing an efficient approach to rapidly screen MRSA directly from clinical specimens has become vital. In this study, we establish a closed-tube loop-mediated isothermal amplification (LAMP) method incorporating hydroxy-naphthol blue (HNB) colorimetric dye assay to directly detect MRSA from clinical samples based on the presence of and genes. In total, 125 preidentified isolates and 93 clinical samples containing were sourced from the microbiology laboratory at Hamad General Hospital (HGH). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were computed based on conventional PCR. The assay demonstrated 100% specificity, 91.23% sensitivity, 0.90 Cohen Kappa (CK), 100% PPV, and 87.8% NPV for the clinical samples, while clinical isolates exhibited 100% specificity, 97% sensitivity, 0.926 CK, 100% PPV, and 88.89% NPV. Compared to cefoxitin disk diffusion, LAMP provided 100% specificity and sensitivity, 1.00 CK, and 100% for PPV and NPV. The study revealed that the closed-tube LAMP incorporating (HNB) dye is a rapid technique with a turnaround time of less than 1 h and high specificity and sensitivity.
抗菌药物耐药性（AMR）的出现，尤其是耐甲氧西林金黄色葡萄球菌（MRSA），对全球健康构成了重大威胁，因为这些细菌对现有的大多数治疗选择越来越耐药。因此，开发一种从临床标本中快速直接筛选MRSA的有效方法变得至关重要。在本研究中，我们建立了一种封闭管环介导等温扩增（LAMP）方法，结合羟基萘酚蓝（HNB）比色染料测定法，基于 基因和 基因的存在直接从临床样本中检测MRSA。总共从哈马德总医院（HGH）微生物实验室获取了125株预先鉴定为 的分离株和93份含有 的临床样本。基于常规PCR计算敏感性、特异性、阳性预测值（PPV）和阴性预测值（NPV）。该检测方法对临床样本的特异性为100%，敏感性为91.23%，科恩kappa系数（CK）为0.90，PPV为100%，NPV为87.8%，而临床分离株的特异性为100%，敏感性为97%，CK为0.926，PPV为100%，NPV为88.89%。与头孢西丁纸片扩散法相比，LAMP的特异性和敏感性均为100%，CK为1.00，PPV和NPV均为100%。研究表明，结合（HNB）染料的封闭管LAMP是一种快速技术，周转时间不到1小时，具有高特异性和敏感性。
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