Journal List > J Bacteriol Virol > v.38(4) > 1033904

TOOLS
Park, Woo, Jung, Lee, Cha, Park, and Lee: Detection of Virulence Genes of Staphyloccus aureus and Staphylococcus epidermidis Isolated from Suprapubic Urine from Infants with Fever
Original Article

J Bacteriol Virol 2008;38(4):189-196.

Published online: 18 January 2008

DOI: https://doi.org/10.4167/jbv.2008.38.4.189

Detection of Virulence Genes of Staphyloccus aureus and Staphylococcus epidermidis Isolated from Suprapubic Urine from Infants with Fever

Hae-Kyung Park1, ∗, So-Youn Woo1, Yun-Jae Jung2, Eun Ok Lee1, Je-Eun Cha1, Hye-Sook Park3, Seung-Joo Lee4

1Department of Microbiology, School of Medicine, Ewha Womans University, Seoul, Republic of Korea

2Deprtment of Microbiology, Graduate School of Medicine, Gachon University of Medicine and Science, Incheon, Republic of Korea

3Department of Preventive Medicine, Ewha Womans University, Seoul, Republic of Korea

4Department of Pediatrics, School of Medicine, Ewha Womans University, Seoul, Republic of Korea

∗Corresponding author: Hae-Kyung Park, MD, PhD. Department of Microbiology, School of Medicine, Ewha Womans University 911-1 Mok-6-Dong, Yang-chun-gu, 158-710, Seoul, Republic of Korea. Phone: +82-2-2650-5737, Fax: +82-2-2653-8891, e-mail: parkhk@ewha.ac.kr

∗∗ This study was supported by Youn Kyung Foundation (2007). June Seng Lee was acknowledged for his skilled cooperation.

      Revised 4 November 20081 December 2008       Accepted 10 December 2008

Copyright © 2008 Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

While methicillin-resistant Staphylococcus aureus (MRSA) isolated from urinary tract infection (UTI) has recently increased in elderly adult urology patients, it has been only rarely reported in infants. Therefore, in this study to understand whether MRSA may be involved in UTI of infants who run fever without other apparent causes, we identified and counted S. aureus and S. epidermidis in suprapubic urine from 750 febrile infants via microbiological methods, and confirmed the counts via multiplex PCR. And we also detected four virulence genes, mecA, PVL, bbp and icaA genes for S. aureus and S. epidermidis via multiplex PCR in the same specimens. S. aureus (28 cases) counts were as follows: >104 CFU/ml (3/28), 102~103 CFU/ml (1/28) and <102~103 CFU/ml (24/28). S. epidermidis (26 cases) counts were as follows: >104 CFU/ml (2/26), 102~103 CFU/ml (4/26) and 102~103 CFU/ml (20/26). S. aureus virulence genes were detected in 26 cases as mecA (16/26, 59.3%), PVL (17/26, 63.0%), bbp (7/26, 26.9%) and icaA (20/26, 76.9%). S. epidermidis virulence genes were detected in 22 cases as mecA (17/22, 81.0%), PVL (15/22, 71.4%), bbp (3/22, 13.6%) and icaA (13/22, 50.1%). Therefore, mecA, PVL and icaA genes of MRSA and MRSE were detected with high positivity in urines from infants with fever. The results demonstrate that community-acquired MRSA or MRSE may be responsible for UTI incidence in febrile infants.

Keywords: Suprapubic urine, Staphylococcus aureus, Staphyococcus epidermidis, mecA, PVL, bbp, icaA

REFERENCES

1). Aires-de-Sousa M., Conceição T., de Lencastre H. Unusually high prevalence of nosocomial Panton-Valentine leukocidin-positive Staphylococcus aureus isolates in Cape Verde Islands. J Clin Microbiol. 44:3790–3793. 2006.
2). Arciola CR., Campoccia D., Gamberini S., Donati ME., Baldassarri L., Montanaro L. Occurrence of ica genes for slime synthesis in a collection of Staphylococcus epidermidis strains from orthopedic prosthesis infections. Acta Orthop Scand. 74:617–621. 2003.
3). Baba-Moussa L., Anani L., Scheftel JM., Couturier M., Riegel P., Haikou N., Hounsou F., Monteil H., Sanni A., Prévost G. Virulence factors produced by strains of Staphylococcus aureus isolated from urinary tract infections. J Hosp Infect. 68:32–38. 2008.
4). Bahrain M., Vasiliades M., Wolff M., Younus F. Five cases of bacterial endocarditis after furunculosis and the ongoing saga of community-acquired methicillin-resistant Staphylococcus aureus infections. Scand J Infect Dis. 38:702–707. 2006.
5). Diep BA., Sensabaugh GF., Somboona NS., Carleton HA., Perdreau-Remington F. Widespread skin and soft-tissue infections due to two methicillin-resistant Staphylococcus aureus strains harboring the genes for Panton-Valentine leucocidin. J Clin Microbiol. 42:2080–2084. 2004.
6). Frebourg NB., Lefebvre S., Baert S., Lemeland JF. PCR-based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. J Clin Microbiol. 38:877–880. 2000.
7). Hoberman A., Chao HP., Keller DM., Hickey R., Davis HW., Ellis D. Prevalence of urinary tract infection in febrile infants. J Pediatr. 123:17–23. 1993.
crossref
8). Issartel B., Tristan A., Lechevallier S., Bruyere F., Lina G., Garin B., Lacassin F., Bes M., Vandenesch F., Etienne J. Frequent carriage of Panton-Valentine leucocidin genes by Staphylococcus aureus isolates from surgically drained abscesses. J Clin Microbiol. 43:3203–3207. 2005.
9). Johnson LB., Venugopal AA., Pawlak J., Saravolatz LD. Emergence of community-associated methicillin-resistant Staphylococcus aureus infection among patients with end-stage renal disease. Infect Control Hosp Epidemiol. 27:1057–1062. 2006.
10). Jones ME., Karlowsky JA., Draghi DC., Thornsberry C., Sahm DF., Bradley JS. Rates of antimicrobial resistance among common bacterial pathogens causing respiratory, blood, urine, and skin and soft tissue infections in pediatric patients. Eur J Clin Microbiol Infect Dis. 23:445–455. 2004.
crossref
11). Kaneko J., Kimura T., Narita S., Tomita T., Kamio Y. Complete nucleotide sequence and molecular characterization of the temperate staphylococcal bacteriophage phiPVL carrying Panton-Valentine leukocidin genes. Gene. 215:57–67. 1998.
12). Karahan ZC., Tekeli A., Adaleti R., Koyuncu E., Dolapci I., Akan OA. Investigation of Panton-Valentine leukocidin genes and SCCmec types in clinical Staphylococcus aureus isolates from Turkey. Microb Drug Resist. 14:203–210. 2008.
13). Lina G., Piémont Y., Godail-Gamot F., Bes M., Peter MO., Gauduchon V., Vandenesch F., Etienne J. Involvement of Panton-Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis. 29:1128–1132. 1999.
14). Manzur A., Dominguez AM., Pujol M., González MP., Limon E., Hornero A., Martín R., Gudiol F., Ariza J. Community-acquired methicillin-resistant Staphylococcus aureus infections: an emerging threat in Spain. Clin Microbiol Infect. 14:377–380. 2008.
15). Martineau F., Picard FJ., Lansac N., Ménard C., Roy PH., Ouellette M., Bergeron MG. Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother. 44:231–238. 2000.
16). Moran GJ., Krishnadasan A., Gorwitz RJ., Fosheim GE., McDougal LK., Carey RB., Talan DA. Methicillin-resistant S. aureus infections among patients in the emergency department. N Engl J Med. 355:666–674. 2006.
17). O'Gara JP. ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiol Lett. 270:179–188. 2007.
18). Osterlund A., Kahlmeter G., Bieber L., Runehagen A., Breider JM. Intrafamilial spread of highly virulent Staphylococcus aureus strains carrying the gene for Panton-Valentine leukocidin. Scand J Infect Dis. 34:763–764. 2002.
19). Otsuka T., Saito K., Dohmae S., Takano T., Higuchi W., Takizawa Y., Okubo T., Iwakura N., Yamamoto T. Key adhesin gene in community-acquired methicillin-resistant Staphylococcus aureus. Biochem Biophys Res Commun. 346:1234–1244. 2006.
20). Papanastasiou DA., Dimitracopoulos G., Drakou A., Haliotis F., Spiliopoulou I. Significant bacteriuria in infants and young children and relation to bacterial species and pyuria. Clin Microbiol Infect. 4:284–287. 1998.
crossref
21). Prévost G., Mourey L., Colin DA., Menestrina G. Staphylococcal pore-forming toxins. Curr Top Microbiol Immunol. 257:53–83. 2001.
crossref
22). Saiman L., O'Keefe M., Graham PL 3rd., Wu F., Saïd-Salim B., Kreiswirth B., LaSala A., Schlievert PM., Della-Latta P. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women. Clin Infect Dis. 37:1313–1319. 2003.
23). Shigemura K., Tanaka K., Okada H., Nakano Y., Kinoshita S., Gotoh A., Arakawa S., Fujisawa M. Pathogen occurrence and antimicrobial susceptibility of urinary tract infection cases during a 20-year period (1983~2002) at a single institution in Japan. Jpn J Infect Dis. 58:303–308. 2005.
24). Supersac G., Prévost G., Piémont Y. Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that staphylococcal leucocidins and gamma-hemolysin are members of a single, two-component family of toxins. Infect Immun. 61:580–587. 1993.
25). Tristan A., Ying L., Bes M., Etienne J., Vandenesch F., Lina G. Use of multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. J Clin Microbiol. 41:4465–4467. 2003.
26). Wang CC., Lo WT., Chu ML., Siu LK. Epidemiological typing of community-acquired methicillin-resistant Staphylococcus aureus isolates from children in Taiwan. Clin Infect Dis. 39:481–487. 2004.
27). Wannet WJ., Spalburg E., Heck ME., Pluister GN., Tiemersma E., Willems RJ., Huijsdens XW., de Neeling AJ., Etienne J. Emergence of virulent methicillin-resistant Staphylococcus aureus strains carrying Panton-Valentine leucocidin genes in The Netherlands. J Clin Microbiol. 43:3341–3345. 2005.
28). Witte W., Strommenger B., Cuny C., Heuck D., Nuebel U. Methicillin-resistant Staphylococcus aureus containing the Panton-Valentine leucocidin gene in Germany in 2005 and 2006. J Antimicrob Chemother. 60:1258–1263. 2007.

Figure 1.
S. aureus and S. epidermidis isolated from the suprapubic urines of infants were confirmed via multiplex PCR. Lane 1 = DNA marker (100 bp ladder); lane 2 = positive control for S. aureus (108 bp); lane 3 = S. aureus positive sample; lane 4 = positive control for S. epidermidis (125 bp); lane 5 = S. epidermidis positive sample; lane 6 = DNA marker; lane 7 = S. epidermidis positive; lane 8 and 9 = S. aureus positive; lane 10 = S. aureus and mecA positive; lane 11 = S. epidermidis and mecA positive; lane 12 = DNA marker.
jbv-38-189f1.tif
Figure 2.
mecA gene and PVL gene were detected via multiplex PCR from S. aureus and S. epidermidis isolated from the suprapubic urines of infants. Lane 1 = positive control for mecA gene (310 bp); lane 2 = positive control for PVL gene (433 bp); lane 3 = mecA positive sample; lane 4 = PVL positive samlple; lane 5 = mecA and PVL positive; lane 6 = DNA marker (100 bp ladder); lane 7 = mecA and PVL positive; lane 8 = PVL positive; lane 9 = mecA and PVL positive; lane 10 = negative control for mecA and PVL gene; lane 11 = mecA and PVL negative sample; lane 12 = DNA marker.
jbv-38-189f2.tif
Figure 3.
bbp gene and icaA gene were detected via multiplex PCR from S. aureus and S. epidermidis isolated from the suprapubic urines of infants. Lane 1 = DNA marker (100 bp ladder); lane 2 = positive control for bbp gene (575 bp); lane 3 = bbp and icaA positive sample; lane 4, 5, and 6 = icaA positive; lane 7 = DNA marker (100 bp ladder); lane 8 = positive control for icaA (103 bp); lane 9 = negative control for icaA; lane 10 = negative control for bbp; lane 11 and 12 = icaA positive; lane 13 = DNA marker.
jbv-38-189f3.tif
Table 1.
Bacterial counts of S. aureus and S. epidermidis in 54 suprapubic urine samples from infants and young children with fever
Group (CFU/ml) S. aureus S. epidermidis
  N % N %
Non-UTI (<102) 24 85.7 20 76.9
Suspicious UTI (102~3) 1 3.6 4 15.4
UTI (≥104) 3 10.7 2 7.7
Total 28 100.0 26 100.0

PM-H = 0.74

Table 2.
Demographic features of the 28 cases in which S. aureus was detected
  UTI (n=3) Suspicious UTI (n=1) Non-UTI (n=24)
Gender (male: female)∗ 0:3 0:1 17:7
Median age (month) 12 3 2
Age group∗∗      
<1 month 0 (0.0) 0 (0.0) 1 (100.0)
1~6 month 0 (0.0) 1 (5.3) 18 (94.7)
6 month ~ 1 years 1 (33.3) 0 (0.0) 2 (66.7)
≥ 1 years 2 (40.0) 0 (0.0) 3 (60.0)

∗ PM-H = 0.0108,

∗∗ PM-H = 0.0125

Table 3.
Demographic features of the 26 cases in which S. epidermidis was detected
  UTI (n=2) Suspicious UTI (n=4) Non-UTI (n=19)
Gender (male: female) 1:1 2:2 10:8
Median age (month) 2 2 4
Age group      
<1 month 0 (0.0) 0 (0.0) 2 (100.0)
1~ 6 month 2 (13.3) 4 (26.7) 9 (60.0)
6 month ~ 1 years 0 (0.0) 0 (0.0) 3 (100.0)
≥ 1 years 0 (0.0) 0 (0.0) 5 (100.0)
Table 4.
Prevalence of virulent genes detected in S. aureus from 26 infants and young children with fever by multiplex PCR
  UTI (n=3) Suspicious UTI (n=1) Non-UTI (n=19) Total (n=26)
mecA (presence) 2 (66.7) 0 (0.0) 14 (60.9) 16 (59.3)
PVL (presence) 2 (66.7) 0 (0.0) 15 (65.2) 17 (63.0)
bbp (presence) 1 (33.3) 0 (0.0) 6 (27.3) 7 (26.9)
icaA (presence) 2 (66.7) 1 (100.0) 17 (77.3) 20 (76.9)
Table 5.
Prevalence of virulent genes detected in S. epidermidis from 22 infants and young children with fever by multiplex PCR
  UTI (n=3) Suspicious UTI (n=1) Non-UTI (n=22) Total (n=26)
mecA∗ (presence)1 0 (0.0) 1 (50.0) 16 (94.1) 17 (81.0)
PVL (presence)1 1 (50.0) 2 (100.0) 12 (70.6) 15 (71.4)
bbp (presence)2 0 (0.0) 0 (0.0) 3 (16.7) 3 (13.6)
icaA (presence)2 1 (100.0) 3 (100.0) 9 (50.0) 13 (50.1)

∗ PM-H = 0.0009

TOOLS
Similar articles