Echovirus 22 is an atypical enterovirus

Coller, Beth Ann; Chapman, Nora M.; Beck, Melinda A.; Pallansch, Mark A.; Gauntt, Charles J.; Tracy, Steven

doi:10.1128/jvi.64.6.2692-2701.1990

Cited by 74 publications

(37 citation statements)

References 80 publications

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“…All enterovirus prototypes were strongly reactive, except for echoviruses 22 and 23, and the positive/ negative ratios (calculated from the negative-control counts) were often 100 or higher. The nonreactivity of echoviruses 22 and 23 was expected, since these viruses are only distantly related to enteroviruses and evidently represent an independent picornavirus group (6,16). In addition, 23 isolates of enteroviruses from the years 1967 to 1992 were tested by the enterovirus PCR-hybridization assay; all were strongly positive (data not shown), indicating that no major changes occurred in this conserved region of the enterovirus genome during this period.…”

Section: Selection Of Primers and Probesmentioning

confidence: 94%

Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization

et al. 1995

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A sensitive method based on PCR followed by liquid-phase hybridization for detection of enterovirus and rhinovirus RNAs in clinical specimens and cell culture supernatants is described. RNA was extracted from stool samples, throat swabs, nasopharyngeal aspirates, cerebrospinal fluid, urine, and plasma with a commercial phenol-guanidinium-chloroform reagent and purified on a polysulfone membrane, on which the reverse transcriptase reaction was also done. Two sets of oligonucleotide primers from the 5 noncoding region of picornaviruses were selected for DNA amplification of 153-bp (enterovirus) and 120-bp (rhinovirus) regions. Double-stranded amplicons were digested into single strands with T7 gene 6 exonuclease and quantitated by an assay using a europium-labeled probe, streptavidin-and biotinylated probe-coated microtitration wells, and time-resolved fluorometry. The sensitivity of the assay was about one template molecule when purified coxsackievirus A9 RNA was used. All enterovirus prototype strains, except echoviruses 22 and 23, and clinical isolates grown in cell culture or suckling mice were strongly positive by the enterovirus PCR-hybridization, as were selected prototype strains and untyped isolates of rhinoviruses by the rhinovirus PCR-hybridization. In a series of 100 clinical specimens tested, the results for 92 agreed with virus culture results. The detection method described will be useful in etiopathogenic studies on enteroviruses and rhinoviruses.Conventional laboratory diagnosis of enterovirus infections includes isolation of the virus in cell culture, less frequently in suckling mice, followed by neutralization typing. The diagnosis can be supported by serology, mainly on the basis of a titer increase between acute-and convalescent-phase serum specimens or detection of immunoglobulin M antibody. Rhinovirus diagnosis is done almost exclusively by culture of the virus followed by acid lability testing. Serology is complicated by more than 100 serotypes. Diagnosis can be delayed because many enteroviruses and rhinoviruses cause cytopathogenic changes in cell cultures only during prolonged incubation, particularly in the first passage. Increased information on enterovirus and rhinovirus sequences (26, 35) has made possible a new and more rapid approach to this diagnostic problem by detection of the viral genome directly in clinical specimens by the PCR. The first application of this technology was for detection of rhinovirus in nasal washes (9, 10). Later, methods were developed to amplify highly conserved sequences present in both viruses followed by a hybridization assay that differentiated between the groups, first with cell culture isolates (15) and then with clinical specimens (25). A number of groups have now reported the use of PCR for direct detection of enteroviruses (1,14,18,24,27,31,33,38,41,43) and rhinoviruses (3,17,18,39) in clinical specimens.In the present report, we describe the development of a highly sensitive nonnested PCR assay followed by liquid-phase hybridization for detection ...

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Section: Selection Of Primers and Probesmentioning

confidence: 94%

Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization

et al. 1995

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“…Viral protein synthesis in infected cell cultures. Labeling of CVB3 proteins was performed as described previously (17). Briefly, HeLa or MFHF cells were plated at 10,000 cells per well in 24-well clusters.…”

Section: Methodsmentioning

confidence: 99%

“…The single long open reading frame is flanked by a 5Ј nontranslated region (NTR), 742 nucleotides (nt) long, and a much shorter 3Ј NTR which terminates in a poly(A) tract (78). Like the polioviruses (PVs) (64; reviewed in reference 72), CVB3 shuts off host cell protein translation in infected HeLa cells (17). The near atomic structure of the CVB3 virion has been solved (7a), demonstrating that the CVB3 capsid is similar in capsid structure to genetically related entero-and rhinoviruses (12,23,31,66).…”

mentioning

confidence: 99%

The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5' nontranslated region

Chapman

Hufnagel

et al. 1995

J Virol

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129

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We report the construction of chimeric coxsackievirus B3 (CVB3) strains in which sequences of an infectious cDNA copy of a noncardiovirulent CVB3 genome were replaced by the homologous sequences from a cardiovirulent CVB3 genome to identify which of 10 predicted genetic sites determine cardiovirulence. Cardiovirulent phenotype expression was consistently linked to nucleotide 234 (U in cardiovirulent CVB3 and C in avirulent CVB3) in the 5 nontranslated region. Reconstructions of the parental noncardiovirulent CVB3 genome from chimeras restored the noncardiovirulent phenotype when tested in mice. Inoculation of severe combined immunodeficient (scid) mice with the noncardiovirulent CVB3 strain resulted in massive cardiomyocyte necrosis in all animals. Sequence analysis of viral genomes isolated from twelve scid mouse hearts showed that only nucleotide position 234 was different (a C3U transition) from that in the input parental noncardiovirulent CVB3 genome. Higher-order RNA structures predicted by two different algorithms did not demonstrate an obvious local effect caused by the C3U change at nucleotide 234. Initial studies of parental and chimeric CVB3 replication in primary cultures of fetal murine heart fibroblasts and in adult murine cardiac myocytes demonstrated that viral RNA transcriptional efficiency is approximately 10-fold lower for noncardiovirulent CVB3 than for cardiovirulent CVB3. CVB3 did not shut off protein synthesis in murine cardiac fibroblasts, nor were levels of viral protein synthesis significantly different as a function of viral phenotype. Taken together, these data support a significant role for determination of the CVB3 cardiovirulence phenotype by nucleotide 234 in the 5 nontranslated region, possibly via a transcriptional mechanism.

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“…E22 and E23 were originally classified as members of the genus Enterovirus (family Picornaviridae) based on classic viral taxonomic characteristics [Wigand and Sabin, 1961]. They possess many properties, however, which distinguish them from other enteroviruses, including distinctive cytopathology and a failure to shut off host protein synthesis [Shaver et al, 1961;Wigand and Sabin, 1961;Coller et al, 1990], and it has been proposed that they be reclassified in a separate genus within the Picornaviridae [Coller et al, 1991;Hyypiä et al, 1992;Stanway et al, 1994]. Of particular importance for molecular diagnostic development, the nucleotide sequences of E22 and E23 are highly diverged from those of conventional enteroviruses [Stanway et al, 1994;Oberste et al, 1998].…”

Section: Introductionmentioning

confidence: 99%

Specific detection of echoviruses 22 and 23 in cell culture supernatants by RT-PCR

1999

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Reverse transcription-polymerase chain reaction (RT-PCR) methods are available for the rapid detection of enteroviruses in clinical specimens or virus isolates. Pan-enterovirus PCR primers, however, fail to amplify echovirus (E) type 22 or 23 because of their extreme sequence divergence from the other enteroviruses. We have developed an RT-PCR method to detect specifically E22 and E23 RNA directly in tissue culture supernatants without a viral RNA purification step. The E22/E23 primers successfully amplified 20 of 20 clinical isolates of E22 and 4 of 4 E23 isolates representing viruses isolated in 15 states over a 19-year period, as well as E22 and E23 prototype strains isolated in the 1950s. The primers did not amplify any of the other 64 enterovirus prototype strains.

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Echovirus 22 is an atypical enterovirus

Cited by 74 publications

References 80 publications

Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization

Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization

The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5' nontranslated region

Specific detection of echoviruses 22 and 23 in cell culture supernatants by RT-PCR

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