Abstract:

The aim was to establish a rapid detection method of expression of the efflux pump genes via determination expression of emrE efflux pump gene from clinical isolated multidrug resistance Shigella spp. strain H24. RNA of multidrug-resistance Shigella H24 was extracted, and real-time fluorescent quantitative RT-PCR was performed to investigate the correlation between emrE gene mRNA expression level and the drug resistance ability. The relative expression content of the efflux pump gene emrE mRNA from Shigella H24 was obtained by real-time fluorescent quantitative RT-PCR, and the relative expression of emrE gene was 20000 times more than house gene gapA. The same sample repeat amplification, and average coefficient of variation of emrE and gapA relative copy numbers was 1.4%和1.3%, it revealed this method detected both relative copy numbers exactly. The high expression of emrE gene could infer that it was related with multidrug resistance of Shigella spp.. This result was consistent with the findings by cloning emrE gene formerly. The SYBR GreenⅠFQ RT-PCR method had a highly sensitivity and specificity, and could be further applied to other efflux drugs, clinical diagnosis and research from multidrug-resistance.