Despite actions to reduce the prevalence of antibiotic resistance, antibiotic resistance remains a major threat in the treatment and control of infections. Therefore, the development of rapid diagnostic methods plays a significant role in the detection of antibiotic resistance genes and the management of hospital infections. This study aimed to design and develop a gold nanoparticle colorimetric probe-based biosensor was developed for faster and more accurate detection of VIM-2 and IMP-1 Metallo-Beta-Lactamases (MBLs) genes in different clinical samples. After identifying 248 clinical isolates of bacteria with standard biochemical methods and evaluation of antibiotic resistance and identifying MBL-producing strains, PCR method was carried out for detecting VIM-2 and IMP-1 genes as a gold standard. Synthesis of AuNPs were done by citrate reduction method and AuNP -IMP-1 biosensor was used for detecting IMP-1 gene, after functionalization of thiol modified oligonucleotides. AuNP biosensor and IMP-1 PCR were compared in terms of detection indices. Method PCR was defined as the gold standard. Bacteria examined, 87 isolates were resistant to carbapenems. Out of the 87 carbapenem-resistant isolates, 85 (34.2%) were phenotypically positive for MBLs. Also, 7 isolates had IMP-1 gene, but none of them carried VIM-2 gene. The AuNP biosensor had 100% sensitivity, specificity, PPV and NPV, respectively. The final detection limit of IMP-1 genomic DNA (LOD) by PCR and AuNP-IMP-1 biosensor technique was 0.1 fg/μl (2.5 fg/25 reactions) and 0.001 fg/μl (0.025 fg/25 reaction), respectively. According to the promising results of Diagnostic indices, AuNP biosensor is a more efficient and accurate method for direct and indirect detection of antibiotic resistance genes in clinical samples.

尽管采取了减少抗生素耐药性发生率的措施，但抗生素耐药性仍然是治疗和控制感染的主要威胁。因此，快速诊断方法的发展对于抗生素耐药基因的检测和医院感染的管理具有重要作用。本研究旨在设计和开发基于金纳米颗粒比色探针的生物传感器，用于更快、更准确地检测不同临床样本中的VIM-2和IMP-1金属β-内酰胺酶 (MBL) 基因。采用标准生化方法对248株临床分离菌进行鉴定并进行抗生素耐药性评价并鉴定产生MBL的菌株后，采用PCR方法检测VIM-2和IMP-1基因作为金标准。采用柠檬酸还原法合成AuNPs，硫醇修饰寡核苷酸功能化后，采用AuNP- IMP-1生物传感器检测IMP-1基因。AuNP生物传感器与IMP-1 PCR在检测指标方面进行了比较。方法 PCR 被定义为金标准。经检查，87 株细菌对碳青霉烯类药物具有耐药性。在 87 株碳青霉烯类耐药菌株中，85 株（34.2%）的 MBL 表型呈阳性。此外，7个分离株具有IMP-1基因，但没有一个分离株携带VIM-2基因。AuNP 生物传感器的灵敏度、特异性、PPV 和 NPV 分别为 100%。IMP-1的最终检出限PCR 和 AuNP- IMP-1生物传感器技术测得的基因组DNA (LOD)分别为 0.1 fg/μl（2.5 fg/25 次反应）和 0.001 fg/μl（0.025 fg/25 次反应）。根据诊断指标的良好结果，AuNP生物传感器是直接和间接检测临床样本中抗生素耐药基因的更有效和准确的方法。