Li Yanan, Feng Yahui, Li Dongmei, Shi Dongmei, Chen Guanzhi
Department of Dermatology, Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.
The Laboratory of Medical Mycology, Jining No. 1 People's Hospital, Jining, Shandong, People's Republic of China.
Infect Drug Resist. 2024 Jul 9;17:2833-2851. doi: 10.2147/IDR.S463798. eCollection 2024.
The increasing incidence of chronic skin infections caused by , coupled with the time-consuming and low detection rates nature of traditional culture and histological-based diagnostic methods, underscores the need for an expedited approach. The study aims to develop a rapid and efficient method for detecting with PCR technology.
We designed four pairs of primers based on DNA sequences from GeneBank and prior studies, we utilized both PCR and Real-time PCR to identify . Specificity and sensitivity assessments were conducted in vitro by DNAs extracted from and other bacterial or fungal cultures. Further validation was performed through the implementation of a mouse skin infection model to optimize and confirm the efficacy of the detection method in both fresh and paraffin-embedded skin tissues. The same PCR testing system was further confirmed with paraffin-embedded skin tissues samples from patients as well.
The results of the study indicate promising outcomes for the four-pair primers system. It demonstrated 100% sensitivity in detecting from purified cultures, including typical strains and nine clinical isolates, while achieving a specificity of 100%. This specificity was evidenced by the absence of PCR products from 12 bacterial species, 12 fungi species, and six other non-tuberculous mycobacterium (NTM) species. In the animal model, the PCR assay exhibited high detection efficacy for both infected fresh tissues and paraffin-embedded tissues, with a slight superiority observed in fresh tissues. However, the PCR assay exhibited high detection efficacy for clinical paraffin-embedded tissues. These findings collectively underscore the robust detection capabilities of our four-pair primers in both in vitro and in vivo settings.
A sensitive and highly specific rapid detection system has been successfully developed that can be used to detect in both infected fresh tissues and paraffin-embedded tissues.
由……引起的慢性皮肤感染发病率不断上升，再加上传统培养和基于组织学的诊断方法耗时且检测率低，凸显了采用快速方法的必要性。本研究旨在开发一种利用PCR技术快速高效检测……的方法。
我们根据基因库中的DNA序列和先前的研究设计了四对引物，利用PCR和实时荧光定量PCR来鉴定……。通过从……以及其他细菌或真菌培养物中提取的DNA进行体外特异性和敏感性评估。通过建立小鼠皮肤感染模型进一步验证，以优化并确认该检测方法在新鲜和石蜡包埋皮肤组织中的有效性。同样的PCR检测系统也用患者的石蜡包埋皮肤组织样本进行了进一步验证。
研究结果表明四对引物系统前景良好。它在检测来自纯化培养物（包括典型菌株和九株临床分离株）中的……时显示出100%的敏感性，同时特异性达到100%。12种细菌、12种真菌和6种其他非结核分枝杆菌（NTM）物种均未产生PCR产物，证明了这种特异性。在动物模型中，PCR检测对感染的新鲜组织和石蜡包埋组织均显示出高检测效能，在新鲜组织中略具优势。然而，PCR检测对临床石蜡包埋组织也显示出高检测效能。这些发现共同强调了我们的四对引物在体外和体内环境中的强大检测能力。
已成功开发出一种灵敏且高度特异的快速检测系统，可用于检测感染的新鲜组织和石蜡包埋组织中的……
