Evaluation of different continuous cell lines in the isolation of mumps virus by the shell vial method from clinical samples.
OBJECTIVE
To compare prospectively the efficacy of the Vero, LLC-MK2, MDCK, Hep-2, and MRC-5 cell lines in the isolation of the mumps virus from clinical samples by means of the shell vial method.


METHODS
During an epidemic outbreak of parotiditis 48 clinical samples (saliva swabs and CSF) were studied. Two vials of the Vero, LLC-MK2, MDCK, MRC-5, and Hep-2 cell lines were inoculated with 0.2 ml of the samples by the shell vial assay. The vials were incubated at 36 degrees C for two and five days. The vials were then fixed with acetone at -20 degrees C for 10 minutes and stained by a monoclonal antibody against mumps virus by means of an indirect immunofluorescence assay.


RESULTS
The mumps virus was isolated from 36 samples. The Vero and LLC-MK2 cell lines showed a 100% isolation capacity, MDCK showed 77.7%, MRC-5 showed 44.4%, and Hep-2 showed 22.2%. The Vero and LLC-MK2 lines were significantly different to the other cell lines (p < 0.001). The sensitivity for the Vero and LLC-MK2 lines at two and five days of incubation was identical (100%). The values obtained in the study of the quantitative isolation capacity (positive isolation with > 5 infectious foci) were 94.4% for Vero, 97.2% for LLC-MK2, 5.5% for MDCK, 5.5% for Hep-2, and 0% for MRC-5.


CONCLUSIONS
The Vero and LLC-MK2 cell lines are equally efficient at two and five days incubation for the isolation of the mumps virus from clinical samples, and the use of the shell vial method considerably shortens the time of aetiological diagnosis with higher specificity.
Reina
J
J
Virology Unit, Clinical Microbiology Service, University Hospital Son Dureta (UIB), Andrea Doria 55, 07014 Palma de Mallorca, Spain. jreina@hsd.es
Ballesteros
F
F
Mari
M
M
Munar
M
M
D000818:Animals; D002460:Cell Line / Q000821:virology; D002522:Chlorocebus aethiops; D006801:Humans; D009107:Mumps / Q000821:virology*; D009109:Mumps virus / Q000302:isolation & purification*; D011446:Prospective Studies; D012463:Saliva / Q000821:virology*; D012680:Sensitivity and Specificity; D014709:Vero Cells / Q000821:virology; D014776:Virus Cultivation / Q000295:instrumentation / Q000379:methods