Impact of cell culture sensitivity and virus concentration on rapid detection of herpes simplex virus by cytopathic effects and immunoperoxidase staining

Abstract

Tremendous interest has been generated in the commercial kits now available that incorporate herpes simplex virus isolation in cell culture with immunoperoxidase staining for viral antigen detection. Most studies comparing commercial kits with conventional cell culture techniques have found the kits to be less sensitive. However, different cell cultures were used for the two methods. In this study, mink lung, rabbit kidney, MRC-5, and Vero cells were compared for reisolation of herpes simplex virus from clinical specimens in which viral infectivity titers were concurrently determined. When specimens contained high titers of infectious virus, the cell system used made little difference and all specimens were detected by immunoperoxidase staining at 48 h postinoculation. However, when specimens contained low concentrations of virus, the differences in sensitivity between cell systems became apparent in rapidity of detection and overall isolation rate. Mink lung and rabbit kidney cells were both more sensitive than MRC-5 cells; Vero cells were significantly less sensitive than the other cells tested. The application of immunoperoxidase staining shortened the time to virus detection and lessened, but did not eliminate, the differences between the cell systems. Cytopathic effects alone in the most sensitive cell system equaled or exceeded immunoperoxidase staining applied in less-sensitive cell cultures.

Comparison of diploid fibroblast and rabbit kidney tissue cultures and a diploid fibroblast microtiter plate system for the isolation of herpes simplex virus

“…In summary, our study indicates that, as others have reported (2,5,10,12), standard tissue cultures using RK cells demonstrated an enhanced sensitivity for HSV. This system should be considered in laboratory situations requiring maximum sensitivity, such as screening for asymptomatic genital shedding of HSV.…”

Conventional tube cell culture compared with centrifugal inoculation of MRC-5 cells and staining with monoclonal antibodies for detection of herpes simplex virus in clinical specimens

“…This is especially important in a reference laboratory, since the time involved in specimen transport from outside the institution results in a delay in processing and, most likely, a fall in virus titer. Cell lines more sensitive to HSV, however, such as mink lung cells (6,9) or possibly A549 cells, may be adequate substitutes in rapid centrifugation methods for conventional cell culture and should be further evaluated. Although good results have been obtained with Vero cells (5), Zhao et al (9) demonstrated that this cell lune was less sensitive than MRC-5 cells to infection with HSV.…”

Detection of sexually transmitted pathogens Trichomonas vaginalis and Herpes simplex virus in women

“…Because the virus is enveloped and highly labile, specimens collected using a swab must be transferred to suitable viral transport media (VTM), such as M4, M6, or universal transport medium, and others [15] . Both rabbit kidney and mink lung cell lines were used for culturing of HSV-1 and HSV-2, with (100% and 95%, respectively) and showed that the sensitivities with a low viral inoculum were higher than those with the MRC-5 and Vero cell lines (77% and 64%, respectively) [17] . Direct immunofluorescence assays (DFA) is another method developed for direct diagnosis of patient specimens which giving the result in one day.…”