1. Comparison of methods for detection of four common nosocomial pathogens on hospital textilesSabina Fijan, Sonja Šostar-Turk, Urška Rozman, 2014, original scientific article Abstract: Introduction: 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.
Keywords: health care associated infections, hospital textiles, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Clostridium difficile, Morapex
Published in DKUM: 05.04.2017; Views: 2185; Downloads: 428
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2. UGOTAVLJANJE VPLIVA PROBIOTIČNIH MIKROORGANIZMOV V KEFIRJU, NA POTENCIALNO PATOGENO BAKTERIJO KLEBSIELLA PNEUMONIAESabina Novak, 2015, undergraduate thesis Abstract: Kefir je z encimi bogato živilo polno telesu koristnih mikroorganizmov, ki ob rednem uživanju obnavljajo našo črevesno floro in povečujejo odpornost. V diplomskem delu smo preučevali probiotike ter kefir, in njegov zaviralni učinek na rast in razmnoževanje bakterije Klebsiella pneumoniae. Opisali smo tudi bakterijo Klebsiella pneumoniae in bakterijsko pljučnico. Diplomsko delo je sestavljeno iz dveh delov; teoretičnega in raziskovalnega. V teoretičnem delu smo opisali probiotike, kefir in njegovo sestavo, kamor sodijo kefirna zrna in kefiran. Predstavljena je bakterija Klebsiella pneumoniae in bolezni ki jih povzroča. V raziskovalnem delu smo preučevali, kako se potencialno patogena bakterija Klebsiella pneumoniae razmnožuje in raste v svežem mleku in ob dodani kulturi kefirnih zrn. Različne koncentracije bakterije Klebsiella pneumoniae smo dodali v sveže mleko ali sveže mleko s kefirnimi zrni in opazovali vpliv probiotičnih mikroorganizmov v kefirju. Na podlagi raziskovalnega dela smo potrdili hipotezo, da prisotnost kefirnih zrn v mleku vpliva na zmanjšanje preživetja bakterije K. pneumoniae. Iz tega lahko sklepamo, da ima kefir antibakterijski učinek.
Keywords: probiotiki, kefir, kefirna zrna, mikrobiota kefirnih zrn, bakterija, Klebsiella pneumoniae
Published in DKUM: 16.02.2016; Views: 2135; Downloads: 206
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3. Real-time polymerase chain reaction for quantitative assessment of common pathogens associated with healthcare-acquired infections on hospital textilesUrška Rozman, Sabina Fijan, Sonja Šostar-Turk, Vid Mlakar, 2013, original scientific article Abstract: 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 frominappropriately disinfected textiles. The aim of the study was to establish qPCR for quantitative assessment of selected common nosocomial pathogens (Clostridium difficile, Staphylococcus aureus, Klebsiella pneumoniaeand 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 presenceof nosocomial pathogens on hospital textiles can be confirmed with qPCR even where conventional techniques do not give any results. qPCR offers apossibility 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.
Keywords: healthcare-acquired infections, hospital textiles, Clostridium difficile, Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa
Published in DKUM: 10.07.2015; Views: 1875; Downloads: 131
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4. Efficiency of four sampling methods used to detect two common nosocomial pathogens on textilesUrška Rozman, Sonja Šostar-Turk, Sabina Fijan, 2012, original scientific article Abstract: Detecting microorganisms on textiles is useful for many purposes, for example to determine the bioburden before laundering, assess the reduction in bacterial counts in connection with various laundry processes, or trace transfer routes in infection control investigations. Therefore a validated, reproducible and rational method is needed. For sampling microorganisms on textile surfaces the most commonly used method is the contact plate method using RODAC plates, first described by Hall and Hartnett followed by the swab sampling technique. Both methods can only capture microorganisms on the surface of the textiles while microorganisms that have penetrated into the deeper structure of the material will not be detected. In our research the contact plate method and the swabbing technique were compared with two wash-off methods. For the first wash-off method the destructive elution methodwas used, where microorganisms were eluted from the fabrics by shaking the fabrics for a certain time in an elution medium. For the fourth sampling method a nondestructive method that included a compact test device called MorapexŽ was used, which is based on forced desorption by pressing the microorganisms through the fabric without destroying the fabric. In our research, two types of microorganisms were included (Klebsiella pneumoniae andStaphylococcus aureus) that cause common nosocomial infections. The aim of this study was to compare the efficiency of the four sampling methods for detecting microorganisms on textiles and to determine the lowest concentration, which can still be detected. The percentage of microorganisms that were detected by both elution methods was substantially higher than by sampling of fabrics with the contact plate method or swabbing. It can be concluded that a nondestructive method using a modified MorapexŽ device can beapplied for quick determination of the hygienic condition of textiles.
Keywords: textile hygiene, Staphylococcus aureus, Klebsiella pneumoniae, swabbing, RODAC plates, elution method, Morapex
Published in DKUM: 10.07.2015; Views: 1701; Downloads: 115
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5. DOLOČEVANJE MEJE DETEKCIJE IZBRANIH MIKROORGANIZMOV NA RAZLIČNIH POVRŠINAH V BOLNIŠNIČNEM OKOLJU Z METODO PCRPatricija Murko, 2015, master's thesis Abstract: Izhodišča: V magistrski nalogi smo določevali mejo detekcije izbranim mikroorganizmom na različnih površinah v bolnišničnem okolju. Prenos s človeka na človeka je najbolj pogost, saj je v bolnišničnem okolju veliko pacientov, prav tako pa tudi zdravstvenega osebja. Različne površine v bolnišničnem okolju lahko tudi pripomorejo k razširitvi mikroorganizmov. Dokazano je, da so mikroorganizmi sposobni preživeti na površinah tudi do nekaj tednov.
Metodologija: V raziskavi smo vzorčili predhodno kontaminirane kovinske, steklene in plastične površine in jemali brise le-teh po 24-urnem sušenju. Izbrani mikroorganizmi so bili: E. faecium, S. aureus, B. subtilis, E. coli, K. pneumoniae, P. aeruginosa in C. albicans. Iz brisov smo eluirali mikroorganizme s fiziološko raztopino. Iz eluata smo najprej določili koncentracijo mikroorganizmov s klasično mikrobiološko metodo z nanosom na selektivne medije. Z metodo PCR smo nato določili še prisotnost DNK izbranih mikroorganizmov. Za statistično analizo smo uporabili programsko orodje IBM SPSS 20. Z neparametričnima testoma smo ugotavljali statistično pomembne razlike.
Rezultati: Z molekularno metodo smo rezultat dobili v nekaj urah, s klasično inkubacijsko metodo je trajalo 2–3 dni. Ko smo primerjali preživetje izbranih mikroorganizmov pri različnih koncentracijah na kovinski in stekleni površini, nismo opazili statistično značilnih razlik, saj je bil p ≥ 0,05. Ko pa smo zajeli še plastično površino, so se razlike pokazale pri po Gramu pozitivnima bakterijama E. faecium in S. aureus ter po Gramu negativnima bakterijama E. coli in P. aeruginosa, saj je bil p ≤ 0,05.
Sklep: Klasična gojitvena metoda je dolgotrajna, medtem ko se metoda PCR lahko uporablja za hitro oceno čistoče površin, saj smo v nekaj urah prišli do rezultatov.
Keywords: bolnišnične okužbe, detekcija, površine, PCR, Enterococcus faecium, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Candida albicans.
Published in DKUM: 15.06.2015; Views: 2455; Downloads: 199
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6. Vzorčenje in dokazovanje prisotnosti klebsiella pneumoniae na bolnišničnih tekstilijah z različnimi metodamiMarko Bočkaj, 2014, undergraduate thesis Abstract: Bolnišnične okužbe so okužbe, povezane z zdravljenjem, bivanjem ali rehabilitacijo v bolnišnici in so najpogostejši zaplet pri zdravljenju. Najpogosteje do njih pride zaradi neupoštevanja in neustreznega izvajanja preventivnih ukrepov za preprečevanje bolnišničnih okužb, med katerimi je najpomembnejša higiena rok. Bolnišnične okužbe povzročajo mikroorganizmi, kot so bakterije, virusi, glive in paraziti. Ena od povzročiteljev takšnih okužb je bakterija Klebsiella pneumoniae, ki je najpomembnejši povzročitelj bolnišničnih okužb iz rodu Klebsiella. Okužbe z bakterijo Klebsiella pneumoniae najpogosteje prizadenejo dihala, sečila in rane. Patogeni mikroorganizmi, ki povzročajo okužbe, se pogosto zadržujejo v tekstilijah, ki se uporabljajo pri vsakdanji praksi v bolnišnici. Zato je detekcija mikroorganizmov v bolnišničnih tekstilijah pomembna pri raziskovanjih na področju obvladovanja in preprečevanja bolnišničnih okužb, ki pestijo bolnike tako dolgo, kot obstajajo bolnišnice in ostale ustanove, ki obravnavajo dlje hospitalizirane bolnike. V naši raziskavi smo primerjali tri različne metode za detekcijo bakterije Klebsiella pneumoniae na bolnišničnih tekstilijah. Najprej smo vzorčili bakterije z klasično metodo jemanja odtisov z RODAC ploščicami, nato z aparatom Morapex A in za konec še z hitrim testom za detekcijo psevdomonad in enterobakterij (Inštitut Hohenstein, Nemčija). Ugotovili smo, da je metoda, pri kateri uporabljamo aparat Morapex A, bolj učinkovita od klasične metode jemanja odtisov z RODAC ploščicami. Hiter test za odkrivanje bakterij na kontaminiranih površinah (Bacteria quick test), pa je neprimeren za uporabo v praksi, saj njegovi rezultati niso tako natančni in zanesljivi kot rezultati, pridobljeni z drugimi metodami vzorčenja.
Keywords: Bolnišnične okužbe, Klebsiella pneumoniae, Morapex A, RODAC, hitri test
Published in DKUM: 22.08.2014; Views: 6695; Downloads: 308
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