Faculty of Medicine, Jan Kochanowski University in Kielce, 25-369 Kielce, Poland.
Dermatology Department, Provincial General Hospital, 25-317 Kielce, Poland.
Int J Mol Sci. 2023 Dec 28;25(1):411. doi: 10.3390/ijms25010411.
The increasing number of patients with chronic wounds requires the development of quick and accurate diagnostics methods. One of the key and challenging aspects of treating ulcers is to control wound infection. Early detection of infection is essential for the application of suitable treatment methods, such as systemic antibiotics or other antimicrobial agents. Clinically, the most frequently used method for detecting microorganisms in wounds is through a swab and culture on appropriate media. This test has major limitations, such as the long bacterial growth time and the selectivity of bacterial growth. This article presents an overview of molecular methods for detecting bacteria in wounds, including real-time polymerase chain reaction (rtPCR), quantitative polymerase chain reaction (qPCR), genotyping, next-generation sequencing (NGS), and loop-mediated isothermal amplification (LAMP). We focus on the LAMP method, which has not yet been widely used to detect bacteria in wounds, but it is an interesting alternative to conventional detection methods. LAMP does not require additional complicated equipment and provides the fastest detection time for microorganisms (approx. 30 min reaction). It also allows the use of many pairs of primers in one reaction and determination of up to 15 organisms in one sample. Isothermal amplification of DNA is currently the easiest and most economical method for microbial detection in wound infection. Direct visualization of the reaction with dyes, along with omitting DNA isolation, has increased the potential use of this method.
慢性伤口患者数量的增加需要开发快速准确的诊断方法。治疗溃疡的一个关键和具有挑战性的方面是控制伤口感染。早期发现感染对于应用合适的治疗方法至关重要,例如全身抗生素或其他抗菌剂。临床上,检测伤口中微生物最常用的方法是通过拭子和适当培养基上的培养。该测试具有主要局限性,例如细菌生长时间长和细菌生长的选择性。本文综述了用于检测伤口中细菌的分子方法,包括实时聚合酶链反应(rtPCR)、定量聚合酶链反应(qPCR)、基因分型、下一代测序(NGS)和环介导等温扩增(LAMP)。我们重点介绍了 LAMP 方法,该方法尚未广泛用于检测伤口中的细菌,但它是对传统检测方法的有趣替代。LAMP 不需要额外复杂的设备,并为微生物提供最快的检测时间(约 30 分钟反应)。它还允许在一个反应中使用多对引物,并在一个样本中确定多达 15 种生物。在伤口感染中,DNA 的等温扩增目前是最简便、最经济的微生物检测方法。用染料直接可视化反应,同时省略 DNA 分离,增加了该方法的潜在用途。