scholarly journals Comparison of methods for detection of four common nosocomial pathogens on hospital textiles

2014 ◽  
Vol 53 (1) ◽  
pp. 17-25 ◽  
Author(s):  
Sabina Fijan ◽  
Sonja Šostar Turk ◽  
Urška Rozman

AbstractIntroduction: 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 phenotype identification. 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 textiles using 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 detectable recovered bacterial concentration on textiles was up to 10 cfu/mL.

1996 ◽  
Vol 76 (03) ◽  
pp. 453-459 ◽  
Author(s):  
Virginie Siguret ◽  
Anne-Sophie Ribba ◽  
Olivier Christophe ◽  
Ghislaine Chérel ◽  
Bernadette Obert ◽  
...  

SummaryThe interacting domain of vWF with platelet GPIb has been shown to overlap the large A1 loop formed by the intra-chain disulfide bond linking Cys 509 to Cys 695. In order to further investigate the role of the conformation of this region, we have expressed in COS-7 cells three mutated full-length recombinant vWFs (rvWFs) in which the substitutions Cys509Gly, Cys509Arg or Cys695Gly have been introduced by site-directed mutagenesis. SDS-agarose gel electrophoresis demonstrated an impaired multimerization of the mutants with undetectable high molecular weight multimers and a decrease of the relative amounts of the intermediate sized multimers. Binding analysis showed that rvWFC509G and rvWFC509R did not interact with botrocetin but spontaneously interacted with GPIb; the latter binding remained unchanged in the presence of ristocetin. This indicates that the substitution of Cys509 by Gly or Arg creates a conformation of vWF that increases its binding to GPIb. In contrast, rvWFC695G which did not react with botrocetin was also unable to interact with GPIb even in the presence of ristocetin, indicating that sequences interacting with GPIb are masked and/or disrupted. In conclusion, the substitution of each of the Cys509 and 695 results in mutant proteins which may be “locked” into active or inactive conformations in regard to the binding to platelet GPIb receptor.


2005 ◽  
Vol 62 (8) ◽  
pp. 1606-1616 ◽  
Author(s):  
John Gilbey ◽  
David Knox ◽  
Martha O'Sullivan ◽  
Eric Verspoor

Abstract Salmon from geographically representative rivers in North America and Europe were typed for variation at the microsatellite locus SS1 and the mitochondrial DNA ND-1 restriction site 3971, using PCR amplification and agarose-gel electrophoresis. North American salmon showed near-fixation for SS1 alleles between 129 and 135 bp in length and the N mtDNA restriction type, while European salmon near-fixation for SS1 alleles between 183 and 219 bp and the A/D mtDNA type. Based on the observed variant frequencies, using these two markers in combination is predicted to give correct assignment of >99.5% of salmon to continent-of-origin. As both these continental markers can be screened by agarose-gel electrophoresis, their use offers a more rapid, cheaper, and simpler method for accurate assignment of Atlantic salmon to continent-of-origin than do existing methods. These markers can be applied to the identification of salmon in North Atlantic high-seas fisheries, in aquaculture stocks, and in rivers to determine the continent-of-origin of fish-farm escapes.


1988 ◽  
Vol 60 (02) ◽  
pp. 133-136 ◽  
Author(s):  
R Schneppenheim ◽  
H Plendl ◽  
U Budde

SummaryA luminescence assay was adapted for detection of von Willebrand factor multimers subsequent to SDS-agarose gel electrophoresis and electroblotting onto nitrocellulose. The method is as fast as chromogenic detection methods and appears to be as sensitive as autoradiography without the disadvantages of the latter.


1969 ◽  
Vol 21 (02) ◽  
pp. 294-303 ◽  
Author(s):  
H Mihara ◽  
T Fujii ◽  
S Okamoto

SummaryBlood was injected into the brains of dogs to produce artificial haematomas, and paraffin injected to produce intracerebral paraffin masses. Cerebrospinal fluid (CSF) and peripheral blood samples were withdrawn at regular intervals and their fibrinolytic activities estimated by the fibrin plate method. Trans-form aminomethylcyclohexane-carboxylic acid (t-AMCHA) was administered to some individuals. Genera] relationships were found between changes in CSF fibrinolytic activity, area of tissue damage and survival time. t-AMCHA was clearly beneficial to those animals given a programme of administration. Tissue activator was extracted from the brain tissue after death or sacrifice for haematoma examination. The possible role of tissue activator in relation to haematoma development, and clinical implications of the results, are discussed.


Author(s):  
M A Jenkins ◽  
M D Guerin

Capillary electrophoresis is a technique that can be automated for the separation of charged particles. By investigating suitable sample dilution and injection time and adhering to a strict washing procedure we have been able to quantify paraproteins in serum samples. This has enabled us to use the technique of capillary electrophoresis for the provision of serum protein electrophoresis in a routine clinical laboratory. We present our findings of 260 serum samples, which included 76 samples with paraproteins analysed by both capillary electrophoresis (EC) and high resolution agarose gel electrophoresis (HRAGE). CE was able to detect all the monoclonal bands detected by HRAGE, and, in particular, better able to detect IgA monoclonal bands occurring in the beta region. The major advantages of CE over HRAGE relate to the automated nature of CE with the elimination of the need for a densitometer.


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