Hatrongjit Rujirat, Chaisaeng Sumontha, Sitthichotthumrong Kulsatree, Boueroy Parichart, Chopjitt Peechanika, Ungcharoen Ratchadaporn, Kerdsin Anusak
Department of General Science, Faculty of Science and Engineering, Kasetsart University Chalermphrakiat, Sakon Nakhon Campus, Sakon Nakhon 47000, Thailand.
Special Research Unit of Emerging Foodborne Pathogens Surveillance, Kasetsart University, Bangkok 10900, Thailand.
Antibiotics (Basel). 2025 Jul 24;14(8):745. doi: 10.3390/antibiotics14080745.
and are common foodborne pathogens of global concern, particularly due to their antimicrobial resistance, notably to cephalosporins. This study aimed to evaluate a polymerase chain reaction-based lateral flow strip (PCR-LFS) assay for the detection of spp. and harboring the gene, which confers resistance to third-generation cephalosporins. Two duplex PCRs (dPCR) were established to detect -harboring (set 1) and -harboring (set 2). 600 bacterial isolates and raw pork mince spiked with -harboring and were used to evaluated. Both dPCR assays successfully detected -positive or strains, while strains lacking the gene showed no amplification. Non- and non- strains were PCR-negative unless they carried . The dPCR-LFS showed 100% validity including accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for both or spp. harboring or lacking . The assay accurately detected target strains without cross-reactivity with other bacteria or antimicrobial resistance genes. Cohen's Kappa coefficient indicated perfect agreement ( = 1), reflecting the high reliability of the dPCR-LFS. The assay could detect as low as 25 CFU/mL for -positive and 40 CFU/mL for -positive in spiked raw pork mince. This assay is rapid, easy to interpret, and suitable for large-scale screening in surveillance programs.
[细菌名称1]和[细菌名称2]是全球关注的常见食源性病原体，特别是由于它们的抗菌耐药性，尤其是对头孢菌素的耐药性。本研究旨在评估一种基于聚合酶链反应的侧向流动试纸条（PCR-LFS）检测方法，用于检测携带赋予对第三代头孢菌素耐药性的[耐药基因名称]基因的[细菌名称1]和[细菌名称2]菌株。建立了两种双重PCR（dPCR）方法来检测携带[耐药基因名称]的[细菌名称1]（组1）和携带[耐药基因名称]的[细菌名称2]（组2）。使用600株细菌分离株以及接种了携带[耐药基因名称]的[细菌名称1]和[细菌名称2]的生猪肉末进行评估。两种dPCR检测方法均成功检测到携带[耐药基因名称]的阳性[细菌名称1]或[细菌名称2]菌株，而缺乏该基因的菌株未出现扩增。非[细菌名称1]和非[细菌名称2]菌株除非携带[耐药基因名称]，否则PCR检测为阴性。dPCR-LFS对携带或不携带[耐药基因名称]的[细菌名称1]或[细菌名称2]菌株的准确性、敏感性、特异性、阳性预测值和阴性预测值均显示为100%有效。该检测方法能准确检测目标菌株，且与其他细菌或抗菌耐药基因无交叉反应。科恩kappa系数表明一致性极佳（κ = 1），反映了dPCR-LFS的高可靠性。该检测方法在接种的生猪肉末中对携带[耐药基因名称]的[细菌名称1]的检测下限低至25 CFU/mL，对携带[耐药基因名称]的[细菌名称2]的检测下限低至40 CFU/mL。该检测方法快速、易于解读，适用于监测项目中的大规模筛查。
