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

Employing a nested polymerase chain reaction with primers from the 5' non-translated region of the enterovirus genome, we detected enteroviral RNA from patients with a variety of enterovirus-associated clinical syndromes. This technique was found to be sensitive (detecting enteroviral RNA extracted from 0.1 50% tissue culture infectious dose) and specific; no specific PCR product was detected from RNA extracts of a variety of non-enterovirus isolates. Although the technique is of comparable sensitivity to single round polymerase chain reaction followed by Southern blot hybridization, it was more rapid, since it enabled a diagnosis to be made within 1 day. Thus, using nested polymerase chain reaction we were able to detect enteroviral RNA in 31 of 46 clinical specimens from 17 of 23 patients with suspected enterovirus infections. The samples included cerebrospinal fluid, throat swabs, stool, vesicle fluid, peripheral blood lymphocytes, whole blood and pericardial effusion. In contrast virus was isolated in only 17 of 42 clinical specimens from 12 of 22 these patients. In preliminary studies, restriction endonuclease analysis of polymerase chain reaction products enabled us to distinguish between non-polio enteroviruses and poliovirus types 1, 2, and 3. This additional technique may be useful in distinguishing between such infections in polio-endemic countries where rapid public health measures may be required.
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PMID:Detection of enterovirus RNA in clinical samples by nested polymerase chain reaction for rapid diagnosis of enterovirus infection. 798 33

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