Author/Editor		Fijan, Sabina; Šostar-Turk, Sonja; Rozman, Urška
Title		Comparison of methods for detection of four common nosocomial pathogens on hospital textiles
Translated title		Primerjava metod za detekcijo štirih pogostih povzročiteljev bolnišničnih okužb na bolnišničnih tekstilijah
Type		članek
Vol. and No.		Letnik 53, št. 1
Publication year		2013
Volume		str. 17-25
Language		eng
Abstract		ntroduction: Although the most common vehicle for transmission of health-care acquired infections is the personto- person transmission route, the role of environment should not be ignored and hospital linen may contribute to the spreading of nosocomial infections. The contact plate method and swabbing are common methods for sampling microorganisms on textiles; however, results are available after two days as they are based on incubation followed by phenotypeidentification. An important alternative is using quick wash-off methods followed by PCR detection, which shortens the identification process from two days to a few hours. Methods: The following test microorganisms at different concentrations were inoculated onto textile swatches and dried overnight: Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Clostridium difficile. RODAC plate sampling as well as a non-destructive wash-off method for capturing microorganisms from the textilesusing a Morapex device were used. The elution suspension from the Morapex device was used for two methods. In the first method, classical incubation on selective media followed by phenotypic identification was used and in the second method DNA was extracted from the elution suspension followed by amplification and agarose gel electrophoresis to visualize amplified products. Conclusions: All chosen bacteria were found using all methods. However, the most sensitive proved to be detection using PCR amplification as we detected the sample with initial concentration of 102 cfu/mL inoculated onto the textile surface before drying. The final detectablerecovered bacterial concentration on textiles was up to 10 cfu/mL.Izhodišča: Čeprav so bolniki ali osebje najpogostejši vir bolnišnično pridobljenih okužb, ne smemo zanemariti vloge okolja, med katere sodijo bolnišnične tekstilije, ki lahko prispevajo k širitvi bolnišničnih okužb. Najpogostejši metodi vzorčenja mikroorganizmov na tekstilijah sta vzorčenje s kontaktnimi ploščami in z brisi, vendar so izsledki na voljo čez dva dni, ker so osnovani na fenotipski identifikaciji po inkubaciji vzorca na gojiščih. Pomembna alternativa so metode eluiranja, ki jim sledi detekcija s PCR-metodo,kar skrajša proces identifikacije z dveh dni na nekaj ur. Metode: Na tekstilne vzorce smo pri različnih koncentracijah nanesli naslednje izbranemikroorganizme - Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa in Clostridium difficile - in jih sušili čez noč. Iz tekstilnih vzorcev smo nato zajeli mikroorganizme z vzorčenjem sploščicami RODAC kakor tudi z nedestruktivno metodo eluiranja z aparatom Morapex. Eluirano suspenzijo iz aparata Morapex smo nadalje uporabili za dve metodi. Pri prvi metodi smo eluat inkubirali na selektivna gojišča, čemur je sledila fenotipska identifikacija, pri drugi metodi pa smo iz eluata ekstrahirali DNK in ga pomnožili ter nato s pomnoženimi produkti izvedli agarozno gelsko elektroforezo. Zaključki: Vse mikroorganizme smo zaznali z vsemi metodami, vendar je bila najobčutljivejša metoda pomnoževanje DNK s PCR-metodo, saj smo po inkubaciji zaznali tudi vzorec z začetno koncentracijo 102 cfu/mL bakterijske suspenzije, inokulirane na tekstilno površino pred sušenjem. Končna koncentracija bakterijske suspenzije, ki smo jo zaznali na tekstilnem vzorcu, je bila do 10 cfu/mL.
Keywords		health care associated infections hospital textiles Staphylococcus aureus Klebsiella pneumoniae Pseudomonas aeruginosa Clostridium difficile Morapex bolnišnično pridobljene okužbe Staphylococcus aureus Klebsiella pneumoniae Pseudomonas aeruginosa Clostridium difficile Morapex