Medical Laboratory Program, Department of Medical Services and Techniques, Vocational School of Health Services, Aksaray University, Aksaray, Turkey.
Department of Medical Microbiology, School of Medicine, Erciyes University, Kayseri, Turkey.
Anaerobe. 2022 Aug;76:102611. doi: 10.1016/j.anaerobe.2022.102611. Epub 2022 Jul 9.
Although anaerobic bacteria are important agents of a wide variety of serious infections, they are overlooked often in the etiology of infection due to difficulties in isolation and detection. The aim of this study was to develop a new multiplex PCR panel that could detect Bacteroides, Fusobacterium, Prevotella, Veillonella, Clostridium, Peptostreptococcus, and Actinomyces bacteria, which are the most frequently isolated from anaerobic infections, at the genus level.
Aerobic and anaerobic cultures were performed on 46 clinical specimens, with suspicion of anaerobic infection and were sent to the laboratory. DNA isolation was performed with the same samples and anaerobic bacteria were detected by the multiplex PCR test developed in the study.
The analytical sensitivity of the multiplex PCR assay was found to be 1-10 CFU/ml, depending on the bacterial species. In this study, anaerobic growth was observed in eight (17.4%) of 46 clinical samples. The multiplex PCR test detected 35 anaerobic bacteria from 20 (43.5%) of 46 clinical samples. The most common anaerobes isolated from clinical specimens by the multiplex PCR assay were Prevotella spp. (37.1%) and Fusobacterium spp. (22.9%) while Clostridium spp. (14.3%), Peptostreptococcus spp. (11.4%), Bacteroides spp. (8.6%), and Veillonella spp. (5.7%) followed these genera.
As a result, it was concluded that the multiplex PCR panel developed in this study eliminates problems in the detection of anaerobes based on culture, provides more accurate detection of anaerobic bacteria from clinical specimens, takes a shorter time, and allows more accurate infection treatment.
尽管厌氧菌是多种严重感染的重要病原体，但由于其分离和检测困难，在感染病因学中常常被忽视。本研究旨在开发一种新的多重 PCR 检测 panel，能够在属水平上检测到从厌氧感染中最常分离到的拟杆菌属、梭杆菌属、普雷沃菌属、韦荣球菌属、梭菌属、消化链球菌属和动弯杆菌属细菌。
对怀疑为厌氧感染并送往实验室的 46 份临床标本进行需氧和厌氧培养。使用相同的样本进行 DNA 分离，并通过本研究中开发的多重 PCR 检测来检测厌氧细菌。
该多重 PCR 检测方法的分析灵敏度为 1-10 CFU/ml，具体取决于细菌种类。在本研究中，46 份临床样本中有 8 份（17.4%）观察到厌氧生长。该多重 PCR 检测从 46 份临床样本中的 20 份（43.5%）中检测到 35 种厌氧细菌。通过多重 PCR 检测从临床标本中分离到的最常见的厌氧菌是普雷沃菌属（37.1%）和梭杆菌属（22.9%），而梭菌属（14.3%）、消化链球菌属（11.4%）、拟杆菌属（8.6%）和韦荣球菌属（5.7%）紧随其后。
因此，本研究开发的多重 PCR 检测 panel 消除了基于培养的厌氧菌检测问题，能够更准确地从临床标本中检测厌氧细菌，检测时间更短，并且能够更准确地进行感染治疗。
