Department of Bacteriology, Capital Institute of Pediatrics, Beijing 100020, China.
Capital Institute of Pediatrics-Peking University Teaching Hospital, Beijing 100020, China.
Anal Chem. 2024 Jun 11;96(23):9424-9429. doi: 10.1021/acs.analchem.4c00450. Epub 2024 Jun 2.
() was first discovered in Japan in 2009 and has since spread worldwide. It exhibits strong transmission ability, high multidrug resistance, blood infectivity, and mortality rates. Traditional diagnostic techniques for have shortcomings, leading to difficulty in its timely diagnosis and identification. Therefore, timely and accurate diagnostic assays for clinical samples are crucial. We developed a novel, rapid recombinase-aided amplification (RAA) assay targeting the 18S rRNA, ITS1, 5.8S rRNA, ITS2, and 28S rRNA genes for identification. This assay can rapidly amplify DNA at 39 °C in 20 min. The analytical sensitivity and specificity were evaluated. From 241 clinical samples collected from pediatric inpatients, none were detected as -positive. We then prepared simulated clinical samples by adding 10-fold serial dilutions of into the samples to test the RAA assay's efficacy and compared it with that of real-time PCR. The assay demonstrated an analytical sensitivity of 10 copies/μL and an analytical specificity of 100%. The lower detection limit of the RAA assay for simulated clinical samples was 10 CFU/mL, which was better than that of real-time PCR (10-10 CFU/mL), demonstrating that the RAA assay may have a better detection efficacy for clinical samples. In summary, the RAA assay has high sensitivity, specificity, and detection efficacy. This assay is a potential new method for detecting , with simple reaction condition requirements, thus helping to manage epidemics.
()于 2009 年在日本首次发现,此后已在全球范围内传播。它具有很强的传播能力、高度的多药耐药性、血液传染性和死亡率。传统的检测技术对于 存在一些缺点,导致难以进行及时的诊断和鉴定。因此,对于临床样本进行及时、准确的诊断检测至关重要。我们开发了一种针对 18S rRNA、ITS1、5.8S rRNA、ITS2 和 28S rRNA 基因的新型快速重组酶辅助扩增(RAA)检测方法,用于 鉴定。该检测方法可在 39°C 下 20 分钟内快速扩增 DNA。我们对该方法的分析灵敏度和特异性进行了评估。从儿科住院患者采集的 241 份临床样本中,均未检测到为 -阳性。然后,我们通过向样本中加入 10 倍系列稀释的 来制备模拟临床样本,以测试 RAA 检测方法的功效,并将其与实时 PCR 进行比较。该检测方法的分析灵敏度为 10 拷贝/μL,分析特异性为 100%。RAA 检测方法对模拟临床样本的最低检测限为 10 CFU/mL,优于实时 PCR(10-10 CFU/mL),表明 RAA 检测方法对临床样本可能具有更好的检测效果。综上所述,该 RAA 检测方法具有高灵敏度、特异性和检测效果。该检测方法是一种用于检测 的潜在新方法,具有简单的反应条件要求,有助于管理 疫情。