Detection of enterovirus RNA in clinical samples by nested polymerase chain reaction for rapid diagnosis of enterovirus infection

Nicholson, Felicity; Meetoo, G; Aiyar, S; Banatvala, J. E.; Muir, Peter

doi:10.1016/0166-0934(94)90115-5

Cited by 66 publications

(48 citation statements)

References 12 publications

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“…Replicate sections of paraffin-embedded formalin-fixed tissue (5-10 ore) were deparaffinized and digested with proteinase K as previously described by Muir et al (1993). Total RNA was extracted from proteinase-K-digested tissue using RNAzol B (Cinna Biotecx), as previously described by Muir et al (1993), and analysed for the presence of enterovirus RNA by reverse transcription followed by nested-PCR amplification (RT-PCR), using primers to amplify a conserved region within the 5' non-translated region of the enterovirus genome (Nicholson et al, 1994). This assay is capable of detecting 0"01 TCIDs0 of enterovirus.…”

Section: Methodsmentioning

confidence: 99%

Enterovirus infection of the central nervous system of humans: lack of association with chronic neurological disease

Muir

Nicholson

Spencer

et al. 1996

Journal of General Virology

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Section: Methodsmentioning

confidence: 99%

Enterovirus infection of the central nervous system of humans: lack of association with chronic neurological disease

Muir

Nicholson

Spencer

et al. 1996

Journal of General Virology

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“…Rapid diagnostic tests have been developed in recent years and are usually based on nucleic acid amplification technology, such as reverse transcription-PCR and nucleic acid sequencebased amplification (3,7,9,12,20,24,31). These assays have proven to be sensitive and specific but, unfortunately, are often in a (semi-)nested format or require specific detection methods, which poses serious hazards for amplification product carryover.…”

mentioning

confidence: 99%

Rapid and Sensitive Routine Detection of All Members of the Genus Enterovirus in Different Clinical Specimens by Real-Time PCR

et al. 2002

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We developed a rapid and sensitive method for the routine detection of all members of the enterovirus genus in different clinical specimens by using real-time TaqMan quantitative PCR. Multiple primer and probe sets were selected in the highly conserved 5-untranslated region of the enterovirus genome. Our assay detected all 60 different enterovirus species tested, whereas no reactivity was observed with the viruses from the other genera of the picornaviridae family, e.g., hepatovirus and parechovirus. Weak cross-reactivity was observed with 7 of the 90 different high-titer rhinovirus stocks but not with rhinovirus-positive clinical isolates. Analysis of a well-characterized reference panel containing different enteroviruses at various concentrations demonstrated that the enterovirus real-time TaqMan PCR is as sensitive as most of the currently used molecular detection assays. Evaluation of clinical isolates demonstrated that the assay is more sensitive than the "gold standard" method, i.e., viral culture. Moreover, the PCR assay can be used on different clinical specimens, such as plasma, serum, nose and throat swabs, cerebrospinal fluid, and bronchoalveolar lavage, without apparent inhibition. Our data demonstrate that the real-time TaqMan PCR is a rapid and sensitive assay for the detection of enterovirus infection. The assay has a robust character and is easily standardized, which makes it an excellent alternative for the conventional time-consuming viral culture.

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“…Most molecular methods are used as diagnostic tools [9,25,27,30,38,46,56,59]. In addition, molecular typing and molecular characterization systems may be used for epidemiological purposes and for the improvement of diagnostic strategies [35].…”

Section: Introductionmentioning

confidence: 99%

Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations

et al. 2012

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Human enteroviruses (HEV) are one of the major causes of central nervous system (CNS) infections in pediatrics. A prospective study was conducted to assess the epidemiological, clinical, and laboratory characteristics of enterovirus (EV) infections of the CNS in children under 15-years-old, suspected of having viral CNS infections and admitted to the Pediatric Department of Monastir University Hospital, Tunisia. Enteroviral RNA was detected by 5 0 NCR nested RT-PCR assay in 33 % (20 out of 60) of cerebrospinal fluid specimens, whereas only six samples (10 %) were EV positive in cell culture. EV-positive patients were clustered according to their clinical manifestations, predominantly diagnosed as aseptic meningitis (65 %) and meningoencephalitis (20 %). Fever, headache, vomiting, and neck stiffness were the most pronounced symptoms. Pleocytosis with the predominance of lymphocytes was observed in 60 % of EV positive specimens. Although patients suffering from EV infections were encountered throughout the year, most occurred during spring and summer months. Using VP1-2A nested RT-PCR and sequence analysis, three of the 20 positive HEV were identified as Echovirus (E)-9. This is the first report of a cluster of aseptic meningitis cases caused by E-9 in Monastir.

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Detection of enterovirus RNA in clinical samples by nested polymerase chain reaction for rapid diagnosis of enterovirus infection

Cited by 66 publications

References 12 publications

Enterovirus infection of the central nervous system of humans: lack of association with chronic neurological disease

Enterovirus infection of the central nervous system of humans: lack of association with chronic neurological disease

Rapid and Sensitive Routine Detection of All Members of the Genus Enterovirus in Different Clinical Specimens by Real-Time PCR

Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations

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