Application of multiplex RT-qPCR in pathogen detection for septic patients

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

Abstract

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

CLC Number:

R631

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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URL: https://www.lchc.cn/EN/10.3969/j.issn.1004-583X.2025.09.004

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

Figures/Tables 4

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

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

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

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

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

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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