Restricted accessResearch articleFirst published online 2013-11
Real-time polymerase chain reaction for quantitative assessment of common pathogens associated with healthcare-acquired infections on hospital textiles
A hospital environment may act as a significant reservoir for potential pathogens that can be transmitted with hospital textiles, which could represent a source of healthcare-acquired infections. Quantitative assessment of nosocomial pathogens with real time polymerase chain reaction (qPCR) on textiles can serve to verify the achievement of standards for textile hygiene of hospital laundry that assess the risk for acquiring hospital infection from inappropriately disinfected textiles. The aim of the study was to establish qPCR for quantitative assessment of selected common nosocomial pathogens (Clostridium difficile, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa) on hospital textiles and to compare the efficiency of the molecular method to the standard procedures for evaluating the bio burden of textiles in hospitals. This study demonstrated that presence of nosocomial pathogens on hospital textiles can be confirmed with qPCR even where conventional techniques do not give any results. qPCR offers a possibility to confirm the presence of microorganisms in dead or viable but non-culturable states that cannot be detected by conventional sampling techniques but may still pose a hazard to public health.
AyliffeGAJCollinsBJLowburyEJL. Ward floors and other surfaces as reservoirs of hospital infection. J Hyg1967; 65: 15–36.
2.
Dancer S, Raeside J, Boothman M. Environmental organisms from different hospital wards. British Journal of Infection Control 2002; 3: 4.
3.
CuaALutwickLI. The environment as a significant cofactor for multiply resistant nosocomial infections. Semin Resp Infect2002; 17: 246–249.
4.
CasewellMPhillipsI. Hands as route of transmission for Klebsiella species. Br Med J1977; 2: 1315–1317.
5.
MariscalALopez-GigososRMCarnero-VaroM. Antimicrobial effect of medical textiles containing bioactive fibres. Eur J Clin Microbiol Infect Dis2011; 30: 227–232.
6.
NeelyAN. A survey of gram-negative bacteria survival on hospital fabrics and plastics. J Burn Care Rehabil2000; 21: 523–527.
7.
BorkowGGabbayJ. Biocidal textiles can help fight nosocomial infections. Med Hypotheses2008; 70: 990–994.
8.
MalnickSBardensteinRHuszarM. Pyjamas and sheets as a potential source of nosocomial pathogens. J Hosp Infect2008; 70: 89–92.
9.
O'HanlonSJEnrightMC. A novel bactericidal fabric coating with potent in vitro activity against meticillin-resistant Staphylococcus aureus (MRSA). Int J Antimicrob Agents2009; 33: 427– 431.
10.
BoyceJMPotter-BynoeGChenevertC. Environmental contamination due to methicillin-resistant Staphylococcus aureus: possible infection control implications. Infect Control Hosp Epidemiol1997; 18: 622–627.
11.
Wiener-WellYGalutyMRudenskyB. Nursing and physician attire as possible source of nosocomial infections. Am J Infect Control2011; 39: 555–559.
12.
PerryCMarshallRJonesE. Bacterial contamination of uniforms. J Hosp Infect2001; 48: 238–241.
13.
AnbazhaganDMuiWSMansorM. Development of conventional and real-time multiplex PCR assays for the detection of nosocomial pathogens. Braz J Microbiol2011; 42: 448–458.
14.
RAL-GZ 992:2001. SACHEGEMËßE WASCHEPFLEGE Gütezicherung RAL-GZ 992 [Proper Linen Care, Quality Assurance RAL-GZ 992]. RAL, Deutsches Institut für Gütezicherung und Kennzeichnung e. V., Sankt Avgustin, p.51.
15.
EN 14065:2003. Textiles - laundry processed textiles - biocontamination control system.
16.
DancerSJ. How do we assess hospital cleaning? A proposal for microbiological standards for surface hygiene in hospitals. J Hosp Infect2004; 56: 10–15.
17.
JIS L 1902:2008. Testing for antibacterial activity and efficacy on textile products. Japanese Standards Association, 4-1–24.
18.
CodyHJSmithPFBlaserMJ. Comparison of methods for recovery of Escherichia coli and Staphylococcus aureus from seeded laundry fabrics. Appl Environ Microbiol1984; 47: 5–5.
19.
EgingtonPJGibsonHHolahJ. Quantification of the ease of removal of bacterial from surfaces. J Ind Microbiol1995; 15: 305–310.
20.
MaunzUKanzE. Beitrage zur quantitativen Auswertung von Abklatsch kulturen. Gesundheitswesen und Desinfektion1969; 61: 129–142.
21.
SuttonS. Accuracy of plate counts. J Validation Technol2011; 17: 3–3.
22.
BabbJRDaviesJGAyliffeGAJ. Contamination of protective clothing and nurses' uniforms in an isolation ward. J Hosp Infect1983; 4: 149–157.
23.
ErikssonBHobornJNyströmB. The contact plate technique is inappropriate for biobured determination on textiles. J Hosp Infect1995; 30: 73–82.
24.
AlsharifRGodfreyW. Bacterial detection and live/dead discrimination by flow cytometry, San Jose, CA: BD Biosciences, 2002.
25.
GonzalezJHIriberriJEgeaL. Characterization of culturability, protistan grazing, and death of enteric bacteria in aquatic ecosystems. Appl Environ Microbiol1992; 58: 998–1004.
26.
BarcinaIGonzalezJMIriberriJ. Effect of visible light on progressive dormancy of Escherichia coli cells during the survival process in natural freshwater. Appl Environ Microbiol1989; 55: 246–251.
27.
BarcinaIGonzalezJMIriberriJ. Survival strategy of Escherichia coli and Enterococcus faecalis in illuminated fresh and marine systems. J Appl Bacteriol1990; 68: 189–198.
28.
GrimesDJAtwellRWBraytonPR. The fate of enteric pathogenic bacteria in estuarine and marine environments. Microbiol Sci1986; 3: 324–329.
29.
MunroPMGauthierMJLaumondFM. Changes in Escherichia coli cells starved in seawater or grown in seawater-wastewater mixtures. Appl Environ Microbiol1987; 53: 1476–1481.
30.
RoszakDBColwellRR. Survival strategies of bacteria in the natural environmental. Microbiol Rev1987; 51: 365– 379.
31.
RoszakDBColwellRR. Metabolic activity of bacterial cells enumerated by direct viable count. Appl Environ Microbiol1987; 53: 2889–2983.
32.
McFetersGAStuartDG. Survival of coliform bacteria in natural waters: field and laboratory studies with membrane-filter chambers. Appl Microbiol1972; 24: 805–811.
33.
KellDBKaprelyantsASWeichartDH. Viability and activity in readily culturable bacteria: a review and discussion of the practical issues. Antonie van Leeuwenhoek1998; 73: 169–187.
34.
ChaiyananSChaiyananSHuqA. Viability of the nonculturable Vibriocholerae O1 and O139. Syst Appl Microbiol2001; 24: 331– 341.
35.
KeerJTBirchL. Molecular methods for the assessment of bacterial viability. J Microbiol Meth2003; 53: 175– 183.
36.
RahmanIShahamatMChowdhuryMAR. Potential virulence of viable but nonculturable Shigella dysenteriae Type1. Appl Environ Microbiol1996; 62: 115–120.
37.
MastersCIShallcrossJAMackeyBM. Effect of stress treatments on the detection of Listeria monocytogenes and enterotoxigenic Escherichia coli by the polymerase chain reaction. J Appl Bacteriol1994; 77: 73–79.
38.
PotéJCeccheriniMTVanVT. Fate and transport of antibiotic resistance genes in saturated soil columns. Eur J Soil Biol2003; 39: 65–71.
39.
SévenoNAKallifidasDSmallaK. Occurrence and reservoirs of antibiotic resistance genes in the environment. Rev Med Microbiol2002; 13: 15–27.
40.
Balamurugan R, Balaji V, Ramakrishna BS. Estimation of faecal carriage of Clostridium difficile in patients with ulcerative colitis using real time polymerase chain reaction. Indian J Med Res 2008; 127: 472–477.
41.
Fusco V, Quero GM, Morea M, Blaiotta G, Visconti A. Rapid and reliable identification of Staphylococcus aureus harbouring the enterotoxin gene cluster (egc) and quantitative detection in raw milk by real time PCR. Int J of Food Microbiology 2011; 144: 528–537.
42.
Liu Y, Liu C, Zheng W, Zhang X, Yu J, Gao Q, Hou Y, Huang X. PCR detection of Klebsiella pneumoniae in infant formula based on 16S–23S internal transcribed spacer. International Journal of Food Microbiology 2008; 125: 230–235.
43.
Motoshimaa M, Yanagiharaa K, Fukushimaa K, Matsudaa J, Sugahara K, Hirakata Y, Yamada Y, Kohno S, Kamihira S. Rapid and accurate detection of Pseudomonas aeruginosa by real-time polymerase chain reaction with melting curve analysis targeting gyrB gene. Diagnostic Microbiology and Infectious Disease 2007; 58: 53–58.
44.
DancerSRaesideJBoothmanM. Environmental organisms from different hospital wards. Br J Infect Control2002; 3: 4–4.
