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1.
Epidemiol Mikrobiol Imunol ; 64(2): 102-6, 2015 Jun.
Article in Czech | MEDLINE | ID: mdl-26099615

ABSTRACT

OBJECTIVE: Introducing enterovirus sequencing as an advanced approach to classify the viruses isolated according to the novel nomenclature and to characterize isolates in detail. MATERIAL AND METHODS: Seventy-five specimens collected from 64 patients in two hospitals, Liberec Regional Hospital, and Plzen University Hospital, were analyzed. The study patients' age ranged from four to 54 years, with a median of 15 years in males and 16 years in females. In most patients, the reasons for admission were intense headache, fever, vomiting, tiredness, meningeal symptoms, intestinal symptoms (in two patients), and skin symptoms (in one patient). The specimens collected were rectal and throat swabs, cerebrospinal fluid (CSF) and stool specimens. Molecular detection and typing were performed using the RT-PCR method. A segment of the 5´non-coding RNA was selected for typing. Specimens were amplified using single-step PCR with external primers and with the same primers extended to include M13 sequences (Generi-Biotech). The LASERGENE software (DIASTAR) was used in sequence editing, alignment, and quality check. The sequences obtained were checked against the central GenBank sequence database using the BLAST algorithm. RESULTS: The identification of the study isolates resulted in 61 ECHO viruses 30, three coxsackie viruses B1, one coxsackie virus B3, one coxsackie virus A9, one enterovirus 86, one enterovirus 71, Two ECHO viruses 13/coxsackie virus B5, one ECHO virus 7/30/coxsackie virus B4, one coxsackie virus B4/enterovirus B, one enterovirus 87/ECHO virus 30/enterovirus B, and one ECHO virus 3. All viruses isolated, except enterovirus 71 classified into group A, were of group B. CONCLUSION: The enteroviruses were identified unambigously, although the sequencing only targeted a short, conserved segment that showed considerable variability. The sequencing was an effective alternative to enterovirus identification by the neutralisation test and allowed for detailed characterization of the isolates. The predominance of ECHO 30 as the cause of aseptic meningitis is in accordance with the literature data.


Subject(s)
Enterovirus Infections/diagnosis , Enterovirus/isolation & purification , Meningitis, Aseptic/diagnosis , Sequence Analysis, DNA/methods , Adolescent , Adult , Child , Child, Preschool , DNA Primers/genetics , Enterovirus/genetics , Enterovirus B, Human/genetics , Enterovirus B, Human/isolation & purification , Enterovirus Infections/virology , Female , Humans , Male , Meningitis, Aseptic/virology , Middle Aged , Neutralization Tests , Polymerase Chain Reaction , Vomiting , Young Adult
2.
Epidemiol Mikrobiol Imunol ; 59(3): 103-6, 2010 Aug.
Article in Czech | MEDLINE | ID: mdl-20925245

ABSTRACT

In the last decade, there has been a rapid development in the use of molecular genetics methods in clinical microbiology. Novel technologies bring new knowledge and approaches to various disciplines of microbiology--taxonomy, identification of microbes, clinical diagnosis, epidemiology of infectious diseases and antibiotic resistance. This article summarizes the conclusions from the workshop of the Molecular Microbiology Working Group TIDE held during the Second Annual Meeting of the Society for Medical Microbiology of the J. E. Purkyne Czech Medical Association.


Subject(s)
Microbiological Techniques , Molecular Biology , Molecular Diagnostic Techniques , Bacteria , DNA, Bacterial/analysis , Humans , Infections/diagnosis
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