Adequate treatment of microbial infections requires rapid and accurate identification of the etiological agent. In routine diagnostics, identification of bacteria conventionally relies on phenotypic testing, which can be hindered by phenotypic variations. Therefore, genotyping techniques should perform faster and more accurately. Recently, the technique of high-resolution melting analysis (HRMA) of PCR amplicons promises to provide a convenient and economic tool of genotypic identification. In our study, we performed prospective routine testing of a PCR-HRMA system that was recently published in a proof-of-the-principle study. The system was evaluated by analysing 275 clinical isolates of bacteria acquired from 65 patients suffering from cystic fibrosis or chronic obstructive pulmonary disease. Our results show that its routine use may result in partial worsening of its discriminatory power; however, it still outmatched conventional phenotyping in the group of non-fermentative Gram-negative rods. Moreover, when supplemented by rapid, simple and economic oxidase test, it can be even simplified for more economic performance.

• MeSH
• Bacteriological Techniques methods   MeSH
• Pulmonary Disease, Chronic Obstructive complications   MeSH
• Cystic Fibrosis complications   MeSH
• Molecular Diagnostic Techniques methods   MeSH
• DNA, Bacterial chemistry   genetics   MeSH
• Gram-Negative Aerobic Rods and Cocci genetics   isolation & purification   MeSH
• Gram-Negative Bacterial Infections diagnosis   microbiology   MeSH
• Respiratory Tract Infections microbiology   MeSH
• Humans MeSH
• Polymerase Chain Reaction methods   MeSH
• Prospective Studies MeSH
• Transition Temperature MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• DNA, Bacterial MeSH

A case report of ventriculoperitoneal shunt infection caused by Candida lusitaniae in a 6-year-old patient with cerebral astrocytoma and obstructive hydrocephalus is presented briefly with emphasis on the course of antifungal treatment. Seven isolates recovered subsequently from the cerebrospinal fluid were studied retrospectively. To confirm identity, isolates were typed using pulsed-field gel electrophoresis and melting curve of random amplified polymorphic DNA (McRAPD). Further, the ability to form biofilm and its susceptibility to systemic antifungals were evaluated. Using McRAPD, identity of C. lusitaniae isolates showing slight microevolutionary changes in karyotypes was undoubtedly confirmed; successful application of numerical interpretation of McRAPD for typing is demonstrated here for the first time. The strain was also recognized as a strong biofilm producer. Moreover, minimum biofilm inhibitory concentrations were very high, in contrast to low antifungal minimum inhibitory concentrations of isolates. It can be concluded that McRAPD seems to be a simple and reliable method not only for identification but also for typing of yeasts. A ventriculoperitoneal shunt colonized by C. lusitaniae was revealed as the source of this nosocomial infection, and the ability of the strain to form biofilm on its surface likely caused treatment failure.

• MeSH
• Antifungal Agents pharmacology   MeSH
• Astrocytoma cerebrospinal fluid   complications   drug therapy   microbiology   pathology   surgery   MeSH
• Biofilms drug effects   growth & development   MeSH
• Candida drug effects   genetics   isolation & purification   MeSH
• Nucleic Acid Denaturation MeSH
• Child MeSH
• Hydrocephalus cerebrospinal fluid   complications   drug therapy   microbiology   pathology   surgery   MeSH
• Cross Infection cerebrospinal fluid   complications   drug therapy   microbiology   pathology   surgery   MeSH
• Candidiasis cerebrospinal fluid   complications   drug therapy   microbiology   pathology   surgery   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Mycological Typing Techniques MeSH
• Brain Neoplasms cerebrospinal fluid   complications   drug therapy   microbiology   pathology   surgery   MeSH
• Treatment Failure MeSH
• Electrophoresis, Gel, Pulsed-Field MeSH
• Random Amplified Polymorphic DNA Technique * MeSH
• Ventriculoperitoneal Shunt adverse effects   MeSH
• Check Tag
• Child MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Case Reports MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antifungal Agents MeSH