VFG-Chip: A high-throughput qPCR microarray for profiling virulence factor genes from the environment

• DOI: 10.1016/j.envint.2023.107761
• Publication Date: 2023-01-16T16:09:54Z
• Authors: Shu-Ting Xie; Long-Jun Ding; Fu-Yi Huang; Yi Zhao; Xin-Li An; Jian-Qiang Su; Guo-Xin Sun; Ya-Qiong Song; Yong-Guan Zhu
• Keywords: Salmonella enterica; Amplicon; Virulence factor

As zoonotic pathogens are threatening public health globally, the virulence factor genes (VFGs) they carry underlie latent risk in environment. However, profiling VFGs environment is still its infancy due to lack of efficient and reliable quantification tools. Here, we developed a novel high-throughput qPCR (HT-qPCR) chip, termed as VFG-Chip, comprehensively quantify abundances targeted A total 96 from four bacterial including Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Salmonella enterica were by 120 primer pairs, which involved encoding five types factors (VFs) like toxin, adherence, secretion system, immune evasion/invasion, iron uptake. The specificity VFG-Chip was both verified computationally experimentally, with high identity amplicon sequencing melting curves analysis proving robust capability. also displayed sensitivity (by plasmid serial dilution test) amplification efficiency averaging 97.7%. We successfully applied profile distribution along wastewater treatment system 69 detected total. Overall, provides tool for quantifying environment, thus information assessing risks