Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.2 (endonuclease)
 18,621 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The aim of this study was to compare conventional enterovirus isolation with rapid detection of enteroviral RNA by a reverse transcription-nested polymerase chain reaction (RT-nPCR) method amplifying the 5' nontranslated region of the enteroviral genome in specimens from patients with aseptic meningitis. Reference enterovirus strains and clinical enterovirus isolates were analyzed to evaluate assay sensitivity and specificity. All known enteroviral serotypes tested, but one (echovirus type 22), were detected by RT-nPCR. A series of unrelated viral isolates as well as CSF samples from patients with meningitis/encephalitis or neurological syndromes unrelated to enterovirus infection were included as controls. A total of 47 specimens (31 CSF, 12 rectal swabs, 4 throat swabs) from 30 patients with aseptic meningitis were available for the study. Of the 31 CSF samples tested from 30 patients, 17 from 17 patients (54.8%) were positive by RT-nPCR, while only 10 from 10 patients (32.2%) were positive by culture. Thus, RT-nPCR allowed diagnosis of enterovirus meningitis in 7 additional patients compared to cell culture. The cytopathic effect was observed 5-15 days after inoculation of CSF specimens onto cell cultures, while direct detection of viral RNA in CSF samples by RT-nPCR permitted diagnosis of enteroviral meningitis within 1-2 days. On the whole, viral isolation was positive in 12/47 (25.5%) specimens, whereas viral RNA was detected by RT-nPCR in 11 additional samples (23/47, 48.9%). Specimens of the control group were consistently negative by both viral isolation and RT-nPCR. Restriction endonuclease analysis of PCR products (RFLP) was applied to differentiate poliovirus (PV) from non-polio enteroviruses (NPEV). All enterovirus strains detected in clinical samples (n = 23) were identified as NPEV by RFLP. Clinical isolates were typed by neutralization as echovirus type 30 (n = 6), while 6 were not typed. In conclusion, detection of enteroviral RNA in CSF by RT-nPCR allows: i) rapid diagnosis of enteroviral meningitis; ii) increased sensitivity with respect to virus isolation; iii) differentiation between PV and NPEV infections of the central nervous system.
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PMID:Rapid detection of enteroviral RNA in cerebrospinal fluid (CSF) from patients with aseptic meningitis by reverse transcription-nested polymerase chain reaction. 981 15

Coxsackie B viruses (genus, Enterovirus; family, Picornaviridae) can cause aseptic meningitis, encephalitis, pleurodynia, myocarditis and are implicated in the pathogenesis of dilated cardiomyopathy. The differentiation of the group B coxsackieviruses into their subtypes has potential clinical and epidemiological implications. In the present study, a simple restriction fragment length polymorphism (RFLP) assay was developed for typing of group B coxsackieviruses into subtypes 1-6. It is a two step process, first, virus isolation and identification by virus neutralization assay, using pools of polyclonal antisera, second, the reverse transcription polymerase chain reaction (RT-PCR) using a single primer pair selected from the conserved 5'-untranslated region (5'-UTR) of enterovirus genome followed by RFLP. A 440 bp product was amplified from the reference strains of each subtype of group B coxsackievirus and 29 clinical isolates (positive for group B coxsackieviruses by neutralization assay). The amplified products were subjected to restriction endonuclease digestion by enzyme BsaJI. The assay was able to distinguish all six serotypes of coxsackie B viruses. The results were comparable to serotyping and showed that due to the relatively conserved nature of 5'-UTR in enterovirus genome, this region can be used for subgeneric molecular identification of enteroviruses.
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PMID:Development of a simple restriction fragment length polymorphism assay for subtyping of coxsackie B viruses. 1528 59