Correlation of carriage and expression of QS regulatory genes and virulence genes with drug resistance in Pseudomonas aeruginosa

Acta Universitatis Medicinalis Anhui 2022 05 v.57 748-753     font:big middle small

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Authors:Zhou Kai; Shi Cuixiao; Song Guobin; Huang Ying; Xu Yuanhong

Keywords:drug resistance;quorum sensing system;virulence genes

DOI:10.19405/j.cnki.issn1000-1492.2022.05.014

〔Abstract〕 Objective To investigate the correlation of carriage and expression of quorum sensing(QS) system regulatory genes and virulence genes with drug resistance inPseudomonas aeruginosastrains isolated from clinical sources. Methods A total of 97 strains ofPseudomonas aeruginosaisolated from various clinical specimens in the laboratory were collected. Firstly, Drug-resistance were detected by the vitek2-compact instrument method and the standard disc diffusion method, 4 QS system-regulated genes and 7 virulence genes were detected by polymerase chain reaction(PCR). After that, the expression of regulatory genes LasI, Rh1 R and virulence genes exoS and PCN were detected by real-time fluorescence quantitative PCR(qRT-PCR), and moreover linearly analysis was used to analyze the correlation of the two types of genes. Results The resistance rates of 97 strains to 11 antibacterial drugs. The resistance rate of tobramycin, gentamicin, and amikacin was less than 10%, and the minimum amikacin was 2.06%. The resistance rate to the remaining drugs was all above 11%, and the highest was imipenem 25.77%, and 43 strains were multi-drug resistant bacteria, and the respiratory-derived strains were more resistant to antimicrobials. The results of PCR gene detection showed: the detection rates of the four QS system-regulated genes LasI, LasR, Rh1 I and Rh1 R were all 100%; the highest detection rate of 7 virulence genes was exoT 98.97%(96/97),and the lowest was exoU 13.40%(13/97),PCN 43.30%(42/97),and the rest were above 86%. The strains without virulence genes were resistant to 6 antibiotics including ceftazidime, and the strains with 7 virulence genes were only resistant to ceftazidime; among the PCN, exoS, and exoU positive strains, the proportion of non-multidrug resistantPseudomonas aeruginosa(NMDR-PA)was higher; PCN+ strains were more resistant to piperacillin/tazobactam and the difference was statistically significant; compared with exoS+/exoU-strains, the types of antibiotics resistant to exoS-/exoU+ strains decreased significantly. The relative expression levels of regulatory genes rh1 R and LasI in NMDR-PAwere higher and the difference of LasI was statistically significant; the expression of the regulatory gene rh1 R and the virulence gene exoS and the regulatory gene LasI and the virulence gene PCN showed a positive linear relationship. Conclusion The drug-resistance ofPseudomonas aeruginosais still serious and multi-drug resistance. QS system regulatory genes exist in the strain stably, except for exoU and PCN, the other virulence genes exoS, exoT, exoY, ToxA and LasB have higher carrying rates, the regulatory genes rh1 R and LasI positively regulate the expression of virulence genes, both QS system regulatory genes and virulence genes affect drug resistance inPseudomonas aeruginosa.