Establishment of a Multiplex Detection Method for Common Bacteria in Blood Based on Human Mannan-Binding Lectin Protein-Conjugated Magnetic Bead Enrichment Combined with Recombinase-Aided PCR Technology

被引:1
作者
Zhao, Zi Jin [1 ,2 ,3 ]
Chen, Xiao Ping [4 ]
Hua, Shao Wei [1 ,2 ,3 ]
Li, Feng Yu [5 ]
Zhao, Meng [2 ,3 ,6 ]
Xing, Chen Hao [1 ]
Wang, Jie [2 ]
Tian, Feng Yu [3 ]
Zhang, Rui Qing [3 ]
Lyu, Xiao Na [2 ,3 ]
Han, Zhi Qiang [2 ,3 ,6 ]
Wang, Yu Xin [2 ,4 ,6 ]
Li, Hong Yi [1 ,2 ,3 ]
Shen, Xin Xin [3 ]
Ma, Xue Jun [3 ]
Tie, Yan Qing [2 ]
机构
[1] Hebei North Univ, Grad Sch, Zhangjiakou 075000, Hebei, Peoples R China
[2] Hebei Gen Hosp, Dept Clin Lab, Shijiazhuang 050051, Hebei, Peoples R China
[3] Chinese Ctr Dis Control & Prevent, Natl Inst Viral Dis Control & Prevent, Natl Key Lab Intelligent Tracking & Forecasting In, NHC Key Lab Med Virol & Viral Dis, Beijing 102206, Peoples R China
[4] Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, Beijing 102206, Peoples R China
[5] Shiyan Gen Hosp, Dept Clin Lab Pathol Diagnost Ctr, Shiyan 442000, Hubei, Peoples R China
[6] Hebei Med Univ, Grad Sch, Shijiazhuang 050031, Hebei, Peoples R China
基金
中国国家自然科学基金;
关键词
Staphylococcus aureus; Pseudomonas aeruginosa; Acinetobacter baumannii; Human Mannan-binding lectin protein; Bloodstream infection; Recombinase-aided PCR assay; Multiple detection; POLYMERASE-CHAIN-REACTION; REAL-TIME PCR; STAPHYLOCOCCUS-AUREUS; PSEUDOMONAS-AERUGINOSA; STREAM INFECTIONS; RAPID DETECTION; CULTURE; ASSAY; MORTALITY; PATHOGENS;
D O I
10.3967/bes2024.043
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
Objective Recombinase-aided polymerase chain reaction (RAP) is a sensitive, single-tube, two-stage nucleic acid amplification method. This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus (SA), Pseudomonas aeruginosa (PA), and Acinetobacter baumannii (AB) in the bloodstream based on recombinant human mannanbinding lectin protein (M1 protein)-conjugated magnetic bead (M1 bead) enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity. Common blood influenza bacteria were used for the specific detection. Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR (M-RAP) and quantitative PCR (qPCR) assays. Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1, 10, and 1 copies/mu L for the detection of SA, PA, and AB plasmids, respectively, without cross-reaction to other bacterial species. The M-RAP assay obtained results for < 10 CFU/mL pathogens in the blood within 4 h, with higher sensitivity than qPCR. M-RAP and qPCR for SA, PA, and AB yielded Kappa values of 0.839, 0.815, and 0.856, respectively (P < 0.05). Conclusion An M-RAP assay for SA, PA, and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.
引用
收藏
页码:387 / 398
页数:12
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