Jiang Luxi, Ren Hongyu, Zhou Haijian, Qin Tian, Chen Yu
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Department of Respiratory Medicine, Shengjing Hospital of China Medical University, Shenyang 110004, China.
Int J Environ Res Public Health. 2017 Feb 23;14(3):223. doi: 10.3390/ijerph14030223.
Early diagnosis and treatment are crucial to the outcome of lower respiratory tract infections (LRTIs). In this study, we developed an assay combining multiplex PCR and Luminex technology (MPLT) for the detection of nine important respiratory bacterial pathogens, which frequently cause LRTIs. These were , , , , , , spp., , and . Through the hybridization reaction between two new synthesized multiplex PCR products and MagPlex-TAG Microspheres, we demonstrate that the detection limits for these nine pathogens were as low as 10²-10³ CFU/mL. Furthermore, 86 clinical bronchoalveolar lavage fluid specimens were used to evaluate this method. Compared with the results of nine simplex real-time PCR reactions targeting these nine pathogens, this MPLT assay demonstrated a high diagnostic accuracy for (sensitivity, 87.5% and specificity, 100%). Furthermore, sensitivity and specificity for the other eight pathogens all attained 100% diagnostic accuracy. In addition, the consistency between MPLT and the nine real-time PCR reactions exceeded 98.8%. In conclusion, MPLT is a high-throughput, labor-saving and reliable method with high sensitivity and specificity for identifying nine respiratory pathogens responsible for LRTIs. Indeed, this assay may be a promising supplement to conventional methods used to diagnose LRTIs.
早期诊断和治疗对下呼吸道感染（LRTIs）的预后至关重要。在本研究中，我们开发了一种结合多重PCR和Luminex技术（MPLT）的检测方法，用于检测九种常见的引起LRTIs的重要呼吸道细菌病原体。它们分别是 、 、 、 、 、 、 属、 、和 。通过两种新合成的多重PCR产物与MagPlex-TAG微球之间的杂交反应，我们证明这九种病原体的检测限低至10²-10³ CFU/mL。此外，使用86份临床支气管肺泡灌洗液体标本来评估该方法。与针对这九种病原体的九种单重实时PCR反应结果相比，这种MPLT检测方法对 显示出较高的诊断准确性（敏感性为87.5%，特异性为100%）。此外，对其他八种病原体的敏感性和特异性均达到100%的诊断准确性。此外，MPLT与九种实时PCR反应之间的一致性超过98.8%。总之，MPLT是一种高通量、省力且可靠的方法，对鉴定导致LRTIs的九种呼吸道病原体具有高敏感性和特异性。实际上，该检测方法可能是诊断LRTIs常用传统方法的一种有前景的补充。
