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Review
. 2007 Jan;20(1):49-78.
doi: 10.1128/CMR.00002-06.

Role of cell culture for virus detection in the age of technology

Diane S Leland  1 , Christine C Ginocchio
Affiliations
Review

Role of cell culture for virus detection in the age of technology

Diane S Leland et al. Clin Microbiol Rev. 2007 Jan.

Abstract

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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Figures

FIG. 1.
FIG. 1.
Uninfected cell cultures and cell cultures showing CPE of viruses commonly isolated. (A) Uninfected A549 cells; (B) HSV-2 in A549 cells; (C) adenovirus in A549 cells; (D) uninfected MRC-5 fibroblasts; (E) CMV in MRC-5 fibroblasts; (F) rhinovirus in MRC-5 fibroblasts; (G) uninfected RhMK cells; (H) enterovirus in RhMk cells; (I) influenza A virus in RhMk cells; (J) uninfected HEp-2 cells; (K) RSV in HEp-2 cells; (L) monkey virus contaminant in RhMk cells. Magnification, ×85.
FIG. 2.
FIG. 2.
Positive hemadsorption result in parainfluenza virus-infected RhMk cells. Magnification, ×100.
FIG. 3.
FIG. 3.
Immunofluorescence detection of respiratory pathogens in R-Mix cells. (A) Uninoculated R-Mix cells; (B) adenovirus; (C) influenza A; (D) influenza B virus; (E) parainfluenza virus type 1; (F) parainfluenza virus type 2; (G) parainfluenza virus type 3; (H) RSV. Magnification, ×170. Photos courtesy of Diagnostic Hybrids, Inc.
FIG. 4.
FIG. 4.
Immunofluorescence detection of Herpesviridae family viruses in H & V cells. (A) Uninoculated H & V cells; (B) CMV; (C) VZV; (D) HSV-1; (E) HSV-2. Magnification, ×170. Photos courtesy of Diagnostic Hybrids, Inc.
FIG. 5.
FIG. 5.
Detection of HSV-1 and HSV-2 in ELVIS cells. (A) Blue cells positive for HSV with X-Gal stain; (B) immunofluorescence of uninoculated ELVIS cells; (C) HSV-1-positive ELVIS immunofluorescence (note nuclear pattern); (D) HSV-2-positive ELVIS immunofluorescence (note cytoplasmic pattern). Magnification, ×170. Photos courtesy of Diagnostic Hybrids, Inc.
FIG. 6.
FIG. 6.
Detection of coxsackievirus B in Super E-Mix cells. (A) Unstained, uninoculated cells; (B) unstained coxsackievirus B CPE; (C and D) immunofluorescence staining with a pan-enterovirus antibody pool of uninoculated Super E-Mix cells (C) and coxsackievirus B-infected cells (D). Magnification, ×170. Photos courtesy of Diagnostic Hybrids, Inc.
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