Isolation of seven respiratory viruses in shell vials: a practical and highly sensitive method

Olsen, Margaret A.; Shuck, Kathleen M.; Sambol, Anthony R.; Flor, Sandra M.; O'Brien, Janet; Cabrera, Barbara J

doi:10.1128/jcm.31.2.422-425.1993

Cited by 44 publications

(14 citation statements)

References 17 publications

(25 reference statements)

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“…By changing the cell line grown on the coverslip and the specificity of the MAbs used for staining, detection of other viruses was easily facilitated. HSV (63), influenza virus (45,130), mumps virus (56), various respiratory viruses (108,117,126), enteroviruses (155), adenoviruses (155), dengue virus (136), and VZV (15,161) have been isolated in shell vials. Recent studies by Landry et al (85) demonstrated the ability to detect hMPV by day 2 postinoculation in A549, HEp-2, and LLC-MK2 shell vials when stained with a MAb (MAb 8510; Chemicon International, Temecula, CA) to hMPV matrix protein.…”

Section: Centrifugation-enhanced Inoculation and Pre-cpe Detection Ofmentioning

confidence: 99%

Role of Cell Culture for Virus Detection in the Age of Technology

2007

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SUMMARY Viral disease diagnosis has traditionally relied on the isolation of viral pathogens in cell cultures. Although this approach is often slow and requires considerable technical expertise, it has been regarded for decades as the “gold standard” for the laboratory diagnosis of viral disease. With the development of nonculture methods for the rapid detection of viral antigens and/or nucleic acids, the usefulness of viral culture has been questioned. This review describes advances in cell culture-based viral diagnostic products and techniques, including the use of newer cell culture formats, cryopreserved cell cultures, centrifugation-enhanced inoculation, precytopathogenic effect detection, cocultivated cell cultures, and transgenic cell lines. All of these contribute to more efficient and less technically demanding viral detection in cell culture. Although most laboratories combine various culture and nonculture approaches to optimize viral disease diagnosis, virus isolation in cell culture remains a useful approach, especially when a viable isolate is needed, if viable and nonviable virus must be differentiated, when infection is not characteristic of any single virus (i.e., when testing for only one virus is not sufficient), and when available culture-based methods can provide a result in a more timely fashion than molecular methods.

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Section: Centrifugation-enhanced Inoculation and Pre-cpe Detection Ofmentioning

confidence: 99%

Role of Cell Culture for Virus Detection in the Age of Technology

2007

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“…The test with the enterovirusspecific MAb scored negative for echovirus types 22, 23, and 25 and some strains of echovirus 1 and 3. Fifteen clinical adenovirus isolates that belonged to types 1,2,3,4,5,7,8,9,10,11,15,17,19,20, and 21 and that had been stored at Ϫ70°C were examined and tested positive by both techniques (data not shown).…”

Section: Influence Of Cell Type On the Rates Of Detection Of Enterovimentioning

confidence: 99%

“…Of the more recently developed methods, the use of nucleic acid amplification techniques for the direct detection of enteroviruses and adenoviruses in clinical specimens is available only in laboratories highly specialized for the diagnosis of viral infections (7). On the other hand, rapid techniques with short-term culture and immunofluorescence for the detection of, for example, respiratory viruses in clinical specimens are widely used (2,6,11,12,15). Application of this approach for the examination of fecal specimens for adenoviruses and enteroviruses has been reported less often (17,19,20).…”

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confidence: 99%

Rapid Shell Vial Culture Technique for Detection of Enteroviruses and Adenoviruses in Fecal Specimens: Comparison with Conventional Virus Isolation Method

Doornum

Jong

1998

J Clin Microbiol

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Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples.

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“…All adenovirus isolates were inoculated in duplicate into human lung carcinoma (A549) cell tube cultures obtained from BioWhittaker Inc. (Walkersville, MD) and ViroMed Laboratories, Inc. (Minnetonka, MN). The use of conventional tube cultures for the isolation of adenoviruses in the clinical setting remains our system of choice, specifically with consideration to those specimens containing low viral titers [21]. All isolates were passaged at least once into tube cultures of the same cell type.…”

Section: Cell Cultures and Specimen Inoculationmentioning

confidence: 99%

“…Utilizing the human lung carcinoma (A549) cell culture as our indicator system, two viral induced monolayer degenerations (i.e., cytopathic effects or CPEs) were recognized. Among our wild and the laboratory adapted (i.e., ATCC) adenovirus isolates tested in this study, serotypes 1,2,4,5,6,8,11,19,21, 27, and 31 were expectedly characterized by the typically enlarged, rounded, and refractile cells, which eventually aggregated into irregular 'grape-like' clusters. Adenovirus types 3 and 7, however, were characterized by the development of distinct intranuclear inclusions, a flattening and then a web or net-like monolayer degeneration.…”

mentioning

confidence: 99%

Presumptive identification of common adenovirus serotypes by the development of differential cytopathic effects in the human lung carcinoma (A549) cell culture

Lipson

Poshni

Ashley

et al. 1993

FEMS Microbiology Letters

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The neutralization test is commonly used in clinical virology laboratories for the identification by serotype of adenovirus isolates. In an effort to conserve reagents and reduce the amount of time in the performance of this assay, we evaluated the significance of differential cytopathic effects for the presumptive identification of lower-numbered adenovirus serotypes that are commonly encountered in the clinical setting. Utilizing the human lung carcinoma (A549) cell culture as our indicator system, two viral induced monolayer degenerations (i.e., cytopathic effects or CPEs) were recognized. Among our wild and the laboratory adapted (i.e., ATCC) adenovirus isolates tested in this study, serotypes 1, 2, 4, 5, 6, 8, 11, 19, 21, 27, and 31 were expectedly characterized by the typically enlarged, rounded, and refractile cells, which eventually aggregated into irregular 'grape-like' clusters. Adenovirus types 3 and 7, however, were characterized by the development of distinct intranuclear inclusions, a flattening and then a web or net-like monolayer degeneration. Differences in the intensity of intranuclear granulation were related by electron microscopy to differences in the quantity of viral crystalline aggregates within the host cell nucleus. A presumptive identification of the commonly encountered adenovirus serotypes 3 and 7 prior to the performance of the neutralization test would result in a conservation of type-specific antiserum, a decreased use of cell cultures and medium, and lastly, reduced medical technologist workload.

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Isolation of seven respiratory viruses in shell vials: a practical and highly sensitive method

Cited by 44 publications

References 17 publications

Role of Cell Culture for Virus Detection in the Age of Technology

Role of Cell Culture for Virus Detection in the Age of Technology

Rapid Shell Vial Culture Technique for Detection of Enteroviruses and Adenoviruses in Fecal Specimens: Comparison with Conventional Virus Isolation Method

Presumptive identification of common adenovirus serotypes by the development of differential cytopathic effects in the human lung carcinoma (A549) cell culture

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