Shinohara Michiyo, Uchida Kazue, Shimada Shin-ichi, Segawa Yukari, Hirose Yoshifumi
Kansenshogaku Zasshi. 2002 Jun;76(6):432-8. doi: 10.11150/kansenshogakuzasshi1970.76.432.
Although various cell lines have been used for virus isolation, few study of virus isolation using MRC-5 cell, a human embryonic lung fibroblasts, have been reported in Japan. MRC-5 and other cell lines (Caco-2, Vero, RD-18s, LLC-MK2, HeLa, MDCK, FL, B95a and HMV-II), and suckling mouse were compared for isolation of viruses from clinical specimens. A total of 3,284 specimens, collected from clinics and hospitals in Saitama Prefecture between July 1997 and August 2001, were inoculated in these cells. A total of 1,252 viral strains were isolated and 1,190 viral strains of these were identified. MRC-5 detected 209 of specimens positive for various viruses. As for adenovirus, a total of 132 viral strains were isolated using cell lines described above, and 100 of 132 viral strains were isolated in MRC-5. MRC-5 showed the highest sensitivity for isolation of adenovirus 3 and 7 (79.1% and 100%) of all other cells. The sensitivity in isolation of these viruses in HeLa was 58.1% and 50.0%, respectively. It showed that MRC-5 is able to isolate enterovirus, especially coxsackie virus A16 and enterovirus 71 with a high sensitivity (85.7% and 73.7%). RD-18s detected 35.7% and 26.3% of coxsackie virus A16 and enterovirus 71 isolates, LLC-MK2 detected 60.7% and 47.4%, and Vero detected 48.6% and 52.6%, respectively. Coxsackie virus B group was not isolated, except for a few coxsackie virus B 5 strains. Enteroviruses except coxsackie virus A16 and enterovirus 71 were isolated more frequently in Caco-2 and RD-18s. Seven hundred thirteen strains of influenza viruses were isolated in MDCK and Caco-2, but none was isolated in MRC-5. It was probably due to the maintenance medium without trypsin. The isolation rate of herpes simplex virus in Vero was 88.9% and MRC-5 showed 77.8%, it was high secondary to Vero by MRC-5. However, the CPE was detected in a few days in MRC-5, it was earlier than in Vero. The MRC-5 is possible to be maintained without changing the maintenance medium and passaged for 2 weeks, and clear CPE was observed. On the other hand, the disadvantages in using the MRC-5 were that the passage was limited and that the split ratio was only 1:2. However, the MRC-5 was used successfully for virus isolation, especially coxsackie virus A16, enterovirus 71 and adenoviruses, from clinical specimens.
尽管已使用多种细胞系进行病毒分离,但在日本,使用人胚肺成纤维细胞MRC-5细胞系进行病毒分离的研究报道较少。比较了MRC-5和其他细胞系(Caco-2、Vero、RD-18s、LLC-MK2、HeLa、MDCK、FL、B95a和HMV-II)以及乳鼠从临床标本中分离病毒的情况。1997年7月至2001年8月期间从埼玉县的诊所和医院收集的总共3284份标本接种于这些细胞。共分离出1252株病毒株,其中1190株病毒株得到鉴定。MRC-5检测出209份各种病毒阳性标本。至于腺病毒,使用上述细胞系共分离出132株病毒株,其中100株在MRC-5中分离得到。MRC-5对腺病毒3型和7型的分离敏感性最高(分别为79.1%和100%),高于所有其他细胞。HeLa对这些病毒的分离敏感性分别为58.1%和50.0%。结果表明,MRC-5能够以高敏感性(分别为85.7%和73.7%)分离肠道病毒,尤其是柯萨奇病毒A16和肠道病毒71。RD-18s检测到柯萨奇病毒A16和肠道病毒71分离株的比例分别为35.7%和26.3%,LLC-MK2检测到的比例分别为60.7%和47.4%,Vero检测到的比例分别为48.6%和52.6%。除少数柯萨奇病毒B5株外,未分离出柯萨奇B组病毒。除柯萨奇病毒A16和肠道病毒71外的肠道病毒在Caco-2和RD-18s中分离更为频繁。在MDCK和Caco-2中分离出713株流感病毒,但在MRC-5中未分离出。这可能是由于维持培养基中没有胰蛋白酶。单纯疱疹病毒在Vero中的分离率为88.9%,MRC-5为77.8%,MRC-5仅次于Vero。然而,MRC-5在几天内就检测到细胞病变效应(CPE),比Vero更早。MRC-5可以在不更换维持培养基的情况下维持并传代2周,且观察到明显的CPE。另一方面,使用MRC-5的缺点是传代有限且分裂比仅为1:2。然而,MRC-5已成功用于从临床标本中分离病毒,尤其是柯萨奇病毒A16、肠道病毒71和腺病毒。