189 Coronaviruses and Toroviruses, Including Severe Acute ...

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2386 SECTION XVII Viral Infectionspossibility that these isolates were of mouse origin has notbeen excluded. In addition, other studies have failed todemonstrate coronavirus RNA in the central nervous systemtissue of patients with multiple sclerosis 99 or have foundsuch RNA in the same proportion of patients with demyelinatingdiseases as in controls. 20 Further investigation will beneeded to establish whether HCoVs are related causally toany neurologic disease in humans.Laboratory DiagnosisVIRUS ISOLATIONRespiratory CoronavirusesIsolation of HCoVs from clinical material has been limited toa few research laboratories. On primary isolation, someHCoV-229E–type strains will replicate and produce a cytopathiceffect in secondary human embryo kidney 31 or certaindiploid cell lines: WI-38, MRC-5, and MA-177. 30, 38 The cellsbecome “stringy” after several days’ incubation in a rollerdrum at 33° C or on subsequent passage. However, someHCoV-229E–like strains require organ culture. 49, 57, 87 In contrast,all HCoV-OC43–like strains seem to require organ culturefor primary isolation, usually human embryonictrachea or nasal mucosa.Enteric Coronavirus-like ParticlesGrowth of these agents in cell or organ culture has beenreported, 15, 88, 115 and one isolate has been adapted to growthin tissue culture. 53TorovirusesHuman toroviruses have not been grown in any culturesystem.VIRUS DETECTION TECHNIQUESRespiratory CoronavirusesRespiratory coronaviruses can be sought successfully inclinical samples by direct antigen-detection techniques orby nucleic acid detection. Some evidence that antigens inHCoV-229E and HCoV-OC43 may be representative of therespiratory HCoV group as a whole exists, 57 although thissuggestion is not accepted universally. Both immunofluorescence68, 95 and ELISA have been used successfully for the35, 56, 69diagnosis of HCoV infection.The HCoV genome has been detected successfully both byRNA-RNA hybridization and by reverse transcription PCR.Hybridization has been reported to detect as little as 10TCID 50 of virus in nasal washes from volunteers infected withHCoV-229E and to have high specificity as well. 77 PCR amplificationof both HCoV-229E and HCoV-OC43 from respiratorysecretions also has been described and applied successfully to76, 82, 83, 93both nasal washings and middle ear fluid. Fewresearchers question that these methods will be useful becausethey are sensitive and specific. In addition, the PCR methodologycan use primers that are specific to the HCoV group, butit may pick up infections caused by strains that might be64, 66related only distantly to the two prototype strains.Enteric CoronavirusesEnteric coronaviruses have been detected primarily by electronmicroscopy of negatively stained preparations fromclarified stool specimens. The possible confusion of coronavirusesand toroviruses in such preparations has been discussedearlier. Immune electron microscopy has beenhelpful in identifying such viruses in purified preparations,but not directly in stool samples.An antigen-detection ELISA for enteric coronavirus hasbeen described and applied to fecal specimens from healthyand diarrheal subjects in Thailand. 51, 52 Findings were similarto those of the electron-microscopic stool studies from India:coronavirus excretion was higher in healthy subjects, andexcretion of viral antigen was observed occasionally to persistfor months. With the adaptation of an antigenically distinctstrain of human enteric coronavirus (HEC) to cell culture,constructing antigen-detection systems for further epidemiologicand clinical study of this virus should be possible. 53TorovirusesAs with enteric coronaviruses, the primary detectionmethod for toroviruses is electron microscopy of clarifiedfecal specimens. Torovirus identity can be specified furtherby either immune electron microscopy or antigen-detectionELISA with the use of antisera prepared against the Bredavirus of calves. 4, 44, 45 With increasing experience, electronmicroscopists can differentiate toroviruses from enteric22, 36coronaviruses on the basis of morphology alone.SERODIAGNOSISFor respiratory coronaviruses, antigens can be preparedwith the use of tissue culture–grown virus or suckling mousebrain preparations. Complement fixation, 17, 63, 67, 71, 90 hemagglutinationinhibition (for HCoV-OC43 only), 42 neutralization,23 ELISA, 13, 28, 46, 56 indirect hemagglutination, 41 andWestern blot 84 all have been used.Serodiagnostic techniques specific for enteric coronaviruseshave not been described. Serodiagnosis oftorovirus infections has been performed both by immuneelectron microscopy and by hemagglutination inhibition. 22Prevention and TreatmentEven if preparing sufficient antigen for vaccine preparationwere possible, the high reinfection rate with these virusessuggests that a vaccine may be ineffective in preventingHCoV-caused respiratory illness. Ribavirin has some in vitroactivity against coronavirus and has been shown to reducehepatitis in mice infected with mouse hepatitis virus, a coronavirusantigenically related to HCoV-OC43. 97AcknowledgmentDr. McIntosh gratefully acknowledges the support of The Bruce R. and JoleneM. McCraw Fund.REFERENCES1. Afzelius, B. A.: Ultrastructure of human nasal epithelium during anepisode of coronavirus infection. Virchows Arch. 424:295–300, 1994.2. Almeida, J. D., and Tyrrell, D. A. J.: The morphology of three previouslyuncharacterized human respiratory viruses that grow in organ culture.J. Gen. Virol. 1:175–178, 1967.3. Arbour, N., Day, R., Newcombe, J., and Talbot, P. J.: Neuroinvasion byhuman respiratory coronaviruses. J. Virol. 74:8913–8921, 2000.4. Beards, G. M., Brown, D. W., Green, J., and Flewett, T. H.: Preliminarycharacterisation of torovirus-like particles of humans: Comparison withBerne virus of horses and Breda virus of calves. J. Med. Virol. 20:67–78,1986.
CHAPTER 189 Coronaviruses and Toroviruses, Including Severe Acute Respiratory Syndrome (SARS) 23875. Beards, G. M., Hall, C., Green, J., et al.: An enveloped virus in stools ofchildren and adults with gastroenteritis that resembles the Breda virus ofcalves. Lancet 1:1050–1052, 1984.6. Bende, M., Barrow, I., Heptonstall, J., et al.: Changes in human nasalmucosa during experimental coronavirus common colds. Acta Otolaryngol.107:262–269, 1989.7. Bradburne, A. F.: Antigenic relationships amongst coronaviruses. Arch.Gesamte Virusforsch. 31:352–364, 1970.8. Bradburne, A. F., Bynoe, M. L., and Tyrrell, D. A.: Effects of a “new”human respiratory virus in volunteers. B. M. J. 3:767–769, 1967.9. Bradburne, A. F., and Somerset, B. A.: Coronavirus antibody titres in seraof healthy adults and experimentally infected volunteers. J. Hyg. (Lond.)70:235–244, 1972.10. Burks, J. S., DeVald, B. L., Jankovsky, L. D., and Gerdes, J. 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