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Koh, Kim, Lee, Jung, Peck, and Lee: The Analysis of the Autoinducer Gene Expression Related Quorum Sensing Mechanism in Catheter Associated Urinary Tract Infection
Original Article
Korean Journal of Urology

Korean Journal of Urology 2006; 47(9): 945-952.

Published online: 30 September 2006

DOI: https://doi.org/10.4111/kju.2006.47.9.945

The Analysis of the Autoinducer Gene Expression Related Quorum Sensing Mechanism in Catheter Associated Urinary Tract Infection

Jun Sung Koh, Hyo Sin Kim, Sang Seob Lee1, Hee Tae Jung2, Kyong Ran Peck3, Ji Youl Lee

Department of Urology, The Catholic University of Korea, Seoul, Korea.

1Department of Biological Engineering, Kyonggi University, Suwon, Korea.

2Department of Chemical Engineering, KAIST, Daejeon, Korea.

3Department of Internal Medicine, Samsung Medical Center, Sungkyunkwan University, Seoul, Korea.

Corresponding author (uroljy@catholic.ac.kr)

Received 4 May 2006       Accepted 1 June 2006

Copyright © 2006 The Korean Urological Association

Abstract

Purpose

Catheter associated urinary tract infection (CAUTI) frequently occurs in the patients with an indwelling Foley catheter, and it can cause serious morbidity or mortality. However, there have been no reports about quorum sensing mechanisms in indwelling Foley catheter. It's our purpose to find out the quorum sensing mechanisms of isolated bacteria from biofilm in Foley catheters.

Materials and Methods

Silicone Foley catheters were placed in 90 patients with neurogenic bladders. At the 3rd, 5th, 7th, 14th and 30th day after the catheters were placed, the catheters were removed and the biofilm formations were evaluated by routine culture and microscopy. The ygaG gene, which was reported to be an autoinducer synthase gene was carried out cloning in E. coli. The quantity of the mRNA expression of the ygaG gene was analyzed according to the time by competitive reverse transcription polymerase chain reaction (RT-PCR).

Results

289 different types of bacteria were isolated by cultivation. The most common species were Pseudomonas, Klebsiella, Serratia, Proteus and Escherichia species. The autoinducer synthase gene, such as the ygaG gene for Escherichia coli, was detected by RT-PCR. On competitive RT-PCR of the ygaG gene, the mRNA expression was 3.77×109copies/µl at the 3rd day, 5.94×107copies/µl at the 5th day, 8.07×107copies/µl at the 7th day, 2.51×106copies/µl at the 14th day and 6.81×108copies/µl at the 30th day. Therefore, the expression of the autoinducer synthase gene was observed at the early insertion period and it was then maintained.

Conclusions

This is the first study to document the autoinducer synthase gene expression associated quorum sensing mechanism in CAUTI. The quorum sensing mechanism may be a new target for the management of CAUTI.

Keywords: Biofilms, Catheter, Antibiotics

Figures and Tables

Fig. 1
(A) The profile of bacteria according to time (in an aerobic condition). (B) The profile of bacteria according to time (in an anaerobic condition).
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Fig. 2
Amplification of the ygaG gene used for RNA sampling by reverse transcription-polymerase chain reaction (RT-PCR). Lane M is the molecular size marker. Lanes 1 and 2 of panels A and B are products of the ygaG gene, which are isolated from bacteria in each of the 3, 5, 7 and 14 days catheters, as the template and ygaG primer. Lane 1 of panel C is the products of the ygaG gene, which is isolated from bacteria in the 30 days catheter.
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Fig. 3
Ethidium bromide stained gel used to generate the equation for quantitative competitive reverse transcription-polymerase chain reaction (RT-PCR) for quantitative analysis of the RNA sample according to the insertion time of the catheter. Lane M is a 1kb DNA ladder. The cDNA of the total RNA from bacteria isolated from a catheter, according to the insertion time, is used as template, and the PCR reactions of lanes 1-9 of panel A contained 10-1, 10-2, 10-3, 10-3.5, 10-3.7, 10-4, 10-3.7, 10-4.1, 10-4.2, respectively, of the competitor cDNA. The PCR reactions of lane 1-6 of panel B contained 10-5, 10-5.3, 10-5.7, 10-2, 10-2.5, 10-3, respectively, of the competitor cDNA.
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Table 1
Oligonucleotide primers for the ygaG and the competitor used for polymerase chain reaction (PCR) amplification and sequencing
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Table 2
The results of the numbers of isolated bacteria from the biofilm of silicons Foley catheters
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Gram: Gram staining, sp: species

Table 3
Quantitative analysis of mRNA expression rate of the ygaG gene according to the insertion time
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