多重RT-qPCR在脓毒症患者病原体检测中的应用

doi:10.3969/j.issn.1004-583X.2025.09.004

摘要/Abstract

摘要：

目的评估多重RT-qPCR在脓毒症患者病原体检测中的临床应用价值。方法选取2024年6月-2025年5月扬州大学第五临床医学院急诊医学科的74例脓毒症患者(SOFA≥2分)及74例健康对照者。检测手段包括血培养(BacT/ALERT^® 3D系统)、多重RT-qPCR(靶向8种常见病原体)及感染标志物(PCT、CRP、IL-6)分析,比较组间差异。结果脓毒症组中,多重RT-qPCR阳性检出率(89.19%)显著高于血培养(75.67%, P=0.027),检测时间显著缩短[(5.42±0.32) h vs (20.25±1.52 ) h,P<0.01]。与血培养相比,多重RT-qPCR敏感度达96.4%,可检出所有血培养阳性样本对应的病原体DNA(κ=0.811)。血培养阳性组及多重RT-qPCR阳性组的感染标志物水平(PCT、CRP、IL-6)均显著高于对照组(P<0.01),但组间差异无统计学意义(P>0.05)。结论多重RT-qPCR具有高敏感度、快速检测及多重靶标覆盖优势,可作为脓毒症早期病原学诊断的可靠补充手段,助力精准抗感染治疗。

关键词: 脓毒症, 多重RT-qPCR, 血培养, 病原体检测, 感染标志物

Abstract:

Objective To evaluate the clinical value of multiplex real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) for pathogen detection in septic patients.Methods A total of 74 septic patients (Sequential Organ Failure Assessment [SOFA]≥2) in the Department of Emergency Medicine of the Fifth Clinical Medical College of Yangzhou University and 74 healthy controls were enrolled from June 2024 to May 2025. Pathogens were detected by blood culture (BacT/ALERT^® 3D), multiplex RT-qPCR (targeting 8 common pathogens), and parallel measurement of infection biomarkers (procalcitonin [PCT], C-reactive protein [CRP], interleukin 6 [IL-6]). Differences between groups were compared. Results The positivity rate of pathogens in septic patients detected by multiplex RT-qPCR was significantly higher than blood culture (89.19% vs 75.67%, P=0.027), with a reduced turnaround time ([5.42±0.32] hours vs [20.25±1.52] hours, P<0.01). Compared to blood culture, multiplex RT-qPCR showed a sensitivity of 96.4% and perfect concordance for positive cases detected by blood culture (κ=0.811). Both blood culture-positive and RT-qPCR-positive groups exhibited significantly elevated infection biomarkers (PCT, CRP, IL-6) compared to controls (P<0.01), but no significant differences were observed between these two groups ( $P$ >0.05). Conclusion Multiplex RT-qPCR demonstrates superior sensitivity, rapidity, and multi-target capacity, serving as a robust adjunct for early etiological diagnosis of sepsis.

Key words: sepsis, multiplex RT-qPCR, blood culture, pathogen detection, infection biomarkers

中图分类号:

R631

沈健, 庾胜, 顾晓蕾, 夏天, 龚菊. 多重RT-qPCR在脓毒症患者病原体检测中的应用[J]. 临床荟萃, 2025, 40(9): 796-800.

Shen Jian, Yu Sheng, Gu Xiaolei, Xia Tian, Gong Ju. Application of multiplex RT-qPCR in pathogen detection for septic patients[J]. Clinical Focus, 2025, 40(9): 796-800.

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链接本文: https://www.lchc.cn/CN/10.3969/j.issn.1004-583X.2025.09.004

https://www.lchc.cn/CN/Y2025/V40/I9/796

图/表 4

表1 血流8种病原体核酸检测试剂盒结果解读

Tab.1 Interpretation of results for nucleic acid test kits for eight bloodstream pathogens

管号	FAM通道	VIC通道	ROX通道	CY5通道
1,6	百日咳杆菌	A族链球菌	/	内标
2,7	肺炎衣原体	/	军团菌	鹦鹉热衣原体
3,8	肺炎科雷伯菌	曲霉菌	新型隐球菌	内标

表2 脓毒症组和对照组的一般资料

Tab.2 General characteristics of the sepsis group and healthy control group

组别	例数	男性[例(%)]	年龄(岁)
脓毒症组	74	33(44.59)	70.03±4.00
对照组	74	38(51.35)	71.01±2.84
$t$ /χ²值		0.677	1.74
$P$ 值		0.411	0.084

表2 脓毒症组和对照组的一般资料

Tab.2 General characteristics of the sepsis group and healthy control group

组别	例数	男性[例(%)]	年龄(岁)
脓毒症组	74	33(44.59)	70.03±4.00
对照组	74	38(51.35)	71.01±2.84
$t$ /χ²值		0.677	1.74
$P$ 值		0.411	0.084

表3 多重RT-qPCR检测效能

Tab.3 Performance of multiplex RT-qPCR detection

多重RT-qPCR	血培养		敏感度	特异度	准确性	阳性预测值	阴性预测值
多重RT-qPCR	阳性(例)	阴性(例)	敏感度	特异度	准确性	阳性预测值	阴性预测值
阳性(例) 阴性(例)	54 2	12 4	0.964	0.333	0.811	0.811	0.750

表4 血培养阳性组、多重 PCR阳性组及对照组间感染性标志物水平比较

Tab.4 Comparison of infectious marker levels among blood culture positive group, multiplex PCR positive group and control group

组别	例数	PCT(ng/L)	CRP(mg/L)	IL-6(pg/L)
血培养阳性组	56	12.50(2.15,38.72)^*	90.25(35.15, 150.30)^*	320.50(50.25, 600.15)^*
多重RT-qPCR阳性组	66	9.75(1.12, 30.64)^*	70.45(40.05, 120.10)^*	400.30(80.10, 850.40)^*
对照组	74	1.05(0.02, 0.12)	4.25(2.05, 8.75)	2.75(0.75, 6.50)
$H$ 值		90.326	95.847	102.614
$P$ 值		<0.01	<0.01	<0.01

表4 血培养阳性组、多重 PCR阳性组及对照组间感染性标志物水平比较

Tab.4 Comparison of infectious marker levels among blood culture positive group, multiplex PCR positive group and control group

组别	例数	PCT(ng/L)	CRP(mg/L)	IL-6(pg/L)
血培养阳性组	56	12.50(2.15,38.72)^*	90.25(35.15, 150.30)^*	320.50(50.25, 600.15)^*
多重RT-qPCR阳性组	66	9.75(1.12, 30.64)^*	70.45(40.05, 120.10)^*	400.30(80.10, 850.40)^*
对照组	74	1.05(0.02, 0.12)	4.25(2.05, 8.75)	2.75(0.75, 6.50)
$H$ 值		90.326	95.847	102.614
$P$ 值		<0.01	<0.01	<0.01

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