Comparative analyses of the detection performance of five multiplex polymerase chain reaction nucleic acid detection kits for respiratory pathogens
10.19428/j.cnki.sjpm.2026.250226
- VernacularTitle:5款呼吸道病原体多重聚合酶链反应核酸检测试剂盒检测性能比对分析
- Author:
Fang YUAN
1
;
Lei BI
2
;
Jiajing LIU
1
;
Huanru WANG
1
;
Jun FENG
1
;
Yuan ZHUANG
1
;
Min CHEN
1
;
Zheng TENG
1
Author Information
1. Institute of Pathogen Biology Identification, Shanghai Municipal Center for Disease Control and Prevention (Shanghai Academy of Preventive Medicine), Shanghai 201107, China
2. Changning District Center for Disease Control and Prevention (Changning District Health Supervision Institute), Shanghai 200335, China
- Publication Type:Journal Article
- Keywords:
respiratory pathogen;
respiratory virus;
multiplex polymerase chain reaction;
nucleic acid detection;
reagent comparison
- From:
Shanghai Journal of Preventive Medicine
2026;38(2):165-169
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo evaluate the detection specificity for clinical samples and the detection capability for standard substances of five commercially available multiplex polymerase chain reaction (PCR) nucleic acid detection kits (hereinafter referred to as the kits) for respiratory pathogens, and to provide a reference for selecting appropriate detection kits for multi-pathogen nucleic acid testing of respiratory infections. MethodsA total of 60 respiratory pathogen-positive clinical samples with known redults were selected and tested using the five kits (labeled as A, B, C, D, and E). The detection rates and Kappa coefficients were calculated to evaluate the consistency between the results from these kits and those from single-pathogen PCR kits. According to the limit of detection (LOD) provided by the kits, standard substances of respiratory pathogens (including 12 types such as influenza virus, Mycoplasma pneumoniae, and Bordetella pertussis) were diluted to four concentrations (250, 500, 1 000, and 2 000 copies·mL⁻¹). All five kits were used for detection to evaluate their respective detection capabilities. ResultsCompared with the results from single-pathogen PCR kits, the five tested kits demonstrated good consistency (all Kappa >0.80). Among them, Kit A had the highest detection rate (100.00%), followed by Kits C and E (98.33%), and then Kits B and D (95.00%). All five kits showed a relatively low false negative rate (FNR) for samples with a cycle threshold (Ct) value ≤35 (≤2.38%). However, for samples with Ct values>35, the FNR increased accordingly(average FNR=6.67%, P=0.029). Kit C exhibited the highest detection sensitivity for the tested standard substances (average LOD: 458.33 copies·mL⁻¹), followed by Kit D, then Kits A/E, and finally Kit B. ConclusionThe five multiplex PCR kits showed good consistency with single-pathogen detection results, but each had its own performance emphasis. Kit A, with the highest detection rate and high throughput, is suitable for targeted viral screening. Kit B, covering the broadest pathogen spectrum (including fungi/bacteria), is suitable for comprehensive respiratory pathogen screening. Kits C, D and E, are applicable for rapid detection. It is important to note that the detection efficacy of all kits decreases for low viral load samples with Ct values >35. In practical application, selection should be based on specific screening objectives, throughput requirements, and sample types.
