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Molecular biological methods in laboratory diagnosis pathogenic leptospires
Kučerová, Petra ; Čermáková, Zuzana (advisor) ; Marešová, Vilma (referee) ; Treml, František (referee)
SUMMMARY Title: "Molecular biological methods in laboratory diagnosis of pathogenic leptospires" During Ph.D. study programe real-time PCR method based on detection of gene encoding surface lipoprotein LipL32 was designed, devised and introduced into clinical laboratory practice of laboratory diagnosis of acute form of leptospirosis. Positive and negative analytical specificity 100 % in both cases was defined and limit of detection in range 1 - 5 copies of genome / ml of liquid biological material was determined. Real-time PCR method was in clinical practice on biological materials gained in period April 2010 - April 2013 from 295 patients suspicious on leptospirosis verified. Total number of 9 persons from whom 15 biological materials originated as LipL32 positive were evaluated. Real-time PCR was tested during the analysis of environmental samples for the presence of Leptospira. From 680 samples (surface water, waste water, wet substrates) were 5 real-time PCR reactions (0.7%) evaluated as borderline, results sequence analysis were repeatedly "uncultivable bacteria." All samples were as negative evaluated. From mention is clear that this method is not suitable for testing these materials. Multilocus sequence typing analysis detecting 5 genes (adk, icdA, rrs2, lipl41 and lipl32 gene) was used to create the...
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