Kok T W, Pryor T, Payne L
Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, Australia.
J Clin Virol. 1998 Jul 24;11(1):61-5. doi: 10.1016/s0928-0197(98)00026-9.
Traditionally, primary monkey kidney (PMK) epithelial cells have been one of the more widely used cell types for the isolation of enteroviruses from clinical samples. For the isolation of coxsackie A viruses, intraperitoneal inoculation of newborn mice has been used in some laboratories.
With the discontinued availability of PMK epithelial cells and the reported growth of coxsackie A viruses in rhabdomyosarcoma cells (RD), we compared the use of the latter cell line with our routinely used microwell cell cultures.
Microwell cell cultures of buffalo green monkey epithelial cell line (BGM), human lung carcinoma epithelial cell line (A549) and human embryonic lung (HEL) fibroblasts were compared with RD cell line for the isolation of enteroviruses from clinical samples.
A total of 39 enteroviruses was isolated from 3517 specimens. The HEL fibroblasts yielded 28 (72%) enteroviruses, followed by A549 (25 isolates, 64%), BGM (23 isolates, 59%) and RD cells (18 isolates, 46%).
All isolates which grew in RD cells showed specific cytopathic effects in one or more of the other inoculated cell cultures. Quantitative determinations (TCID50) with five different enteroviruses showed that the HEL fibroblasts and RD cell line to be the most sensitive cell types, followed by BGM and A549 cell lines. However, integrity of the inoculated cell monolayers was best with HEL fibroblasts and A549 cells but morphology was not optimal with RD cells during the incubation period of 14 days.
传统上,原代猴肾(PMK)上皮细胞一直是从临床样本中分离肠道病毒时使用较为广泛的细胞类型之一。在一些实验室中,对于柯萨奇A病毒的分离,采用了新生小鼠腹腔接种法。
鉴于PMK上皮细胞已停止供应,且有报道称柯萨奇A病毒可在横纹肌肉瘤细胞(RD)中生长,我们将后一种细胞系与我们常规使用的微孔细胞培养法进行了比较。
将水牛绿猴上皮细胞系(BGM)、人肺癌上皮细胞系(A549)和人胚肺(HEL)成纤维细胞的微孔细胞培养法与RD细胞系进行比较,用于从临床样本中分离肠道病毒。
从3517份标本中共分离出39株肠道病毒。HEL成纤维细胞分离出28株(72%)肠道病毒,其次是A549细胞(25株,64%)、BGM细胞(23株,59%)和RD细胞(18株,46%)。
在RD细胞中生长的所有分离株在一种或多种其他接种的细胞培养物中均表现出特异性细胞病变效应。对五种不同肠道病毒进行的定量测定(半数组织培养感染剂量)表明,HEL成纤维细胞和RD细胞系是最敏感的细胞类型,其次是BGM和A549细胞系。然而,在14天的培养期内,接种的细胞单层完整性在HEL成纤维细胞和A549细胞中最佳,但RD细胞的形态并不理想。