通过空斑试验法从临床样本中分离腮腺炎病毒时不同连续细胞系的评估。

Evaluation of different continuous cell lines in the isolation of mumps virus by the shell vial method from clinical samples.

作者信息

Reina J, Ballesteros F, Mari M, Munar M

机构信息

Virology Unit, Clinical Microbiology Service, University Hospital Son Dureta (UIB), Andrea Doria 55, 07014 Palma de Mallorca, Spain.

出版信息

J Clin Pathol. 2001 Dec;54(12):924-6. doi: 10.1136/jcp.54.12.924.

DOI:10.1136/jcp.54.12.924

PMID:11729211

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1731329/

Abstract

AIMS

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.

摘要

目的

前瞻性比较Vero、LLC-MK2、MDCK、Hep-2和MRC-5细胞系通过空斑试验从临床样本中分离腮腺炎病毒的效果。

方法

在腮腺炎流行爆发期间，对48份临床样本（唾液拭子和脑脊液）进行研究。通过空斑试验，将0.2 ml样本接种到两瓶Vero、LLC-MK2、MDCK、MRC-5和Hep-2细胞系中。将培养瓶在36℃孵育2天和5天。然后将培养瓶在-20℃用丙酮固定10分钟，并通过间接免疫荧光试验用抗腮腺炎病毒单克隆抗体进行染色。

结果

从36份样本中分离出腮腺炎病毒。Vero和LLC-MK2细胞系的分离能力为100%，MDCK为77.7%，MRC-5为44.4%，Hep-2为22.2%。Vero和LLC-MK2细胞系与其他细胞系有显著差异（p<0.001）。Vero和LLC-MK2细胞系在孵育2天和5天时的敏感性相同（100%）。在定量分离能力研究中（阳性分离且感染灶>5个）得到的值为：Vero为94.4%，LLC-MK2为97.2%，MDCK为5.5%，Hep-2为5.5%，MRC-5为0%。