Rapid Detection of Pathogens and Resistance Genes Grown in Blood Cultures with Two Multiplex Tandem Real-Time PCR Kits.

Infectious diseases & clinical microbiology Pub Date : 2025-03-27 eCollection Date: 2025-03-01 DOI:10.36519/idcm.2025.465
Barış Gülhan, Aytekin Çıkman, Merve Aydın, Mürşit Hasbek, Tuncer Özekinci, Sümeyye Akyüz, Faruk Karakeçili
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Abstract

Objective: In this study, two multiplex tandem real-time PCR kits were used to rapidly diagnose common Gram-positive cocci and Gram-negative bacilli, detect their commonly seen antibiotic resistance genes, and evaluate the two kits' performance.

Materials and methods: Gram-positive 12 (GP-12) kit (AusDiagnostics, Australia) and Gram-negative 12 (GN-12) kit (AusDiagnostics, Australia) were used in the study. Seventy-eight Gram-negative bacilli and 54 Gram-positive cocci grown in blood culture vials were applied to GN-12 and GP-12 panels. At the same time, the passages of the samples were made and incubated. After that, identification and antibiograms were made in the Phoenix™ automated system (Becton, Dickinson and Company, USA) and VITEK 2 Compact automated system (bioMérieux, France).

Results: Twenty-one Staphylococcus aureus, twelve coagulase-negative staphylococci (CoNS), two Streptococcus pneumoniae, two Enterococcus faecium, and three Enterococcus faecalis were found to match the results from the automated GP-12 Kit. Pathogens present in the panel were successfully identified using the GN-12 kit. Both panels were found to be more effective in diagnosing polymicrobial infections.

Conclusion: These evaluated kits were rapid (approximately three hours) and valuable in identifying common sepsis pathogens and resistance genes. Thus, these tests can easily be used in the diagnosis of sepsis.

用两种多重串联实时荧光定量PCR试剂盒快速检测血培养物中病原菌和抗性基因。
目的:采用两种多重串联实时PCR试剂盒快速诊断常见革兰氏阳性球菌和革兰氏阴性杆菌,检测其常见的抗生素耐药基因,并对两种试剂盒的性能进行评价。材料和方法:采用革兰氏阳性12 (GP-12)试剂盒(AusDiagnostics, Australia)和革兰氏阴性12 (GN-12)试剂盒(AusDiagnostics, Australia)。将血培养瓶中培养的78株革兰氏阴性杆菌和54株革兰氏阳性球菌分别应用于GN-12和GP-12面板。同时,制作样品的传代并孵育。之后,在Phoenix™自动化系统(Becton, Dickinson and Company, USA)和VITEK 2 Compact自动化系统(biomacrieux, France)中进行鉴定和抗生素图谱。结果:共检出21株金黄色葡萄球菌、12株凝固酶阴性葡萄球菌、2株肺炎链球菌、2株粪肠球菌和3株粪肠球菌,与全自动GP-12试剂盒检测结果吻合。使用GN-12试剂盒成功地鉴定了面板中存在的病原体。两种检测方法在诊断多微生物感染方面都更为有效。结论:这些评估试剂盒快速(约3小时),在鉴定常见败血症病原体和耐药基因方面具有重要价值。因此,这些测试可以很容易地用于败血症的诊断。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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