Application of mixed cell lines for the detection of viruses from clinical specimens

Huang, Yung T.; Hite, Scott; Duane, Visa; Yam, Paul; Jollick, Joseph A.; Goodrum, Gail R.

doi:10.1016/s0196-4399(00)89072-8

Cited by 6 publications

(1 citation statement)

References 9 publications

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“…In this study, we compare two sensitive cell culture systems, ELVIS cells for the rapid detection of herpes simplex virus (HSV) (5, 7) and R-Mix (mixture of A549 and mink lung) cells for the detection of influenza viruses A and B (1,2,3,4,6), in both frozen and nonfrozen monolayer formats, to determine whether frozen monolayers can match the virus detection performance of fresh, commercially prepared monolayers.…”

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Cryopreserved Cell Monolayers for Rapid Detection of Herpes Simplex Virus and Influenza Virus

et al. 2002

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Cryopreserved cell monolayers are a new cell culture technology intended to ensure the availability of cells in the laboratory for virus detection. Two cryopreserved cell monolayers, ELVIS for the detection of herpes simplex virus (HSV) and R-Mix for the detection of influenza virus, were evaluated. The results indicated that fresh and cryopreserved cell monolayers are comparable in sensitivity for the detection of HSV and influenza virus. The cells retain the same level of sensitivity for up to 4 months at ؊80°C.A diagnostic virology laboratory generally receives fresh cells from commercial sources once or twice a week. Receipts of commercially prepared cells can be compromised by shipping delays, mishandling of packages, and exposure to temperature stress, which may lead to suboptimal performance. Additionally, because these shipments are estimated standing orders, shortage or overage is common. Finally, the quality of cell monolayers is variable from lot to lot and difficult to control and standardize. These fundamental shortcomings of commercially prepared cells are generally accepted by clinical virology laboratories because the only functional alternative, i.e., preparing one's own cells each week, is generally impractical for technical and/or economic reasons.Recently, Diagnostic Hybrids Inc. (DHI, Athens, Ohio) developed a cryopreservation method that addresses the practical issues cited above. In this study, we compare two sensitive cell culture systems, ELVIS cells for the rapid detection of herpes simplex virus (HSV) (5, 7) and R-Mix (mixture of A549 and mink lung) cells for the detection of influenza viruses A and B (1, 2, 3, 4, 6), in both frozen and nonfrozen monolayer formats, to determine whether frozen monolayers can match the virus detection performance of fresh, commercially prepared monolayers.The cryopreserved ELVIS and R-Mix cell monolayers (ready cell frozen monolayers [RCFM]) were provided on a glass coverslip in shell vials by DHI. RCFM were shipped on dry ice and transferred quickly to storage in a Ϫ80°C chest freezer. The nonfrozen cell equivalents were commercially produced and shipped by express courier, as is routinely done. Prior to inoculation with clinical specimens, cryopreserved ELVIS RCFM and R-Mix RCFM vials were removed from the Ϫ80°C freezer and placed in an empty 24-well cluster plate. The plate was gently placed in a 35 to 37°C water bath such that the water level was just high enough to flood the plate. The vials were incubated for 4 min (Ϯ15 s) without any agitation. The thawed vials were gently removed from the water bath, and the freeze medium was immediately removed from the vials by gentle aspiration.For ELVIS RCFM, 1 ml of ELVIS replacement medium (DHI) was added to each vial and 0.2 ml of clinical specimen was inoculated into both RCFM and fresh cells. All shell vials were centrifuged at 700 ϫ g for 60 min at room temperature and incubated at 35 to 37°C for 20 to 24 h. ELVIS cells were fixed and stained for HSV detection and typing by using the ELVIS HSV ID/typing...

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