【摘要】 目的 系统比较靶向测序(tNGS)、多重实时荧光定量PCR(多重qPCR)与传统病原学检测(CMTs)在呼吸道感染病原体诊断中的性能。方法 回顾性分析2023年1月至2024年12月济源市疾病预防控制中心收集的1 980例呼吸道感染患者样本,所有样本均同步进行tNGS、多重qPCR及CMTs检测。以CMTs结果为参照,计算并比较三种方法的阳性检出率、混合感染检出率、罕见病原体检出率和检测耗时,并采用Kappa检验评估其与CMTs的一致性。结果 1 980份样本中,CMTs的阳性检出率为45.15%,多重qPCR为48.73%,tNGS为59.15%,三者比较差异有统计学意义(P<0.05),而两两比较tNGS的阳性检出率明显高于多重qPCR和CMTs,差异有统计学意义(P<0.05)。tNGS的混合感染检出率和罕见病原体检出率分别为25.70%、12.30%,亦明显高于多重qPCR的18.30%、3.80%和CMTs的10.61%、2.63%,差异均有统计学意义(P<0.05)。多重qPCR检测耗时最短为(2.51±1.49) h,显著短于tNGS的(36.35±11.21) h,差异有显著统计学意义(P<0.01)。在与CMTs的一致性方面,tNGS的Kappa值为0.89(95%CI:0.85～0.92),明显高于多重qPCR的0.76(95%CI:0.71～0.81,P<0.05)。结论 tNGS在呼吸道病原体检测中展现出更高的灵敏度和广谱性,尤其在识别混合感染与罕见病原体方面优势明显,但其检测周期较长。多重qPCR则以其快速、操作简便见长。两种新技术相较于CMTs均能提升检出能力,在临床应用中可形成互补,为精准抗感染治疗提供更全面的病原学依据。更多还原

【Abstract】 Objective To systematically compare the performance of targeted sequencing(tNGS), multiplex real-time fluorescence quantitative PCR(multiplex qPCR), and conventional microbiological tests(CMTs) in the diagnosis of respiratory tract infections. Methods A retrospective analysis was conducted on 1 980 samples from patients with respiratory tract infections collected by Jiyuan Center for Disease Control and Prevention from January 2023 to December 2024. All samples were simultaneously tested by tNGS, multiplex qPCR, and CMTs. Using CMTs as the reference, the positive detection rate, mixed infection detection rate, rare pathogen detection rate, and turnaround time were calculated and compared among the three methods. Consistency with CMTs was evaluated using the Kappa test. Results Among the 1 980 samples, the positive detection rates were 45.15% for CMTs, 48.73% for multiplex qPCR, and 59.15% for tNGS, with a statistically significant difference among the three(P<0.05). Pairwise comparison showed that the positive detection rate of tNGS was significantly higher than those of multiplex qPCR and CMTs(P<0.05). The detection rates of mixed infection and rare pathogens by tNGS were 25.70% and 12.30%, respectively, which were significantly higher than 18.30% and 3.80% by multiplex qPCR and 10.61% and 2.63% by CMTs(all P<0.05). The turnaround time of multiplex qPCR was the shortest at(2.51±1.49) hours, which was significantly shorter than(36.35±11.21) hours of tNGS(P<0.01). Regarding consistency with CMTs, the Kappa value of tNGS was 0.89(95% CI: 0.85-0.92), which was significantly higher than 0.76(95% CI: 0.71-0.81) of multiplex qPCR(P<0.05). Conclusion tNGS demonstrates higher sensitivity and broader spectrum in respiratory pathogen detection, with particular advantages in identifying mixed infections and rare pathogens, but its turnaround time is longer. Multiplex qPCR is characterized by speed and ease of operation.Compared with CMTs, both new technologies improve detection capability and can complement each other in clinical practice, providing a more comprehensive etiological basis for precise anti-infective therapy.更多还原