CACO-2 细胞：一种连续细胞系，对呼吸道病毒培养具有敏感和广谱的应用价值。

CACO-2 cells: A continuous cell line with sensitive and broad-spectrum utility for respiratory virus culture.

机构信息

Laboratory of Viral Diseases, Wadsworth Center, Albany, NY, USA.

出版信息

J Virol Methods. 2021 Jul;293:114120. doi: 10.1016/j.jviromet.2021.114120. Epub 2021 Mar 4.

DOI:10.1016/j.jviromet.2021.114120

PMID:33676967

Abstract

BACKGROUND

Primary rhesus monkey kidney cells (RhMK) can be used for the detection of respiratory viruses, including influenza and parainfluenza. The human colon adeno-carcinoma cell line, CACO-2, has been previously used for the growth of multiple influenza viruses, including seasonal, novel and avian lineages.

OBJECTIVE

We compared CACO-2, Madin-Darby Canine Kidney (MDCK), and RhMK cells for the isolation of viruses from patients presenting with influenza like-illness (ILI).

STUDY DESIGN

Nasopharyngeal specimens from patients with ILI in primary care settings were processed for conventional viral culture in MDCK, RhMK, and CACO-2. Cells were examined microscopically for cytopathic effect (CPE) and confirmatory testing included immunofluorescent antigen (IFA) detection and real-time RT-PCR. Additionally, 16 specimens positive for respiratory syncytial virus (RSV) by PCR were inoculated on CACO-2 cells. Statistical analysis was done using Chi-square test with IBM Statistical Program.

RESULTS

Of 1031 respiratory specimens inoculated, viruses were isolated and confirmed from 331 (32.1 %) in MDCK cells, 304 (29.5 %) in RhMk cells, and 433 (42.0 %) in CACO-2 cells. These included influenza A/(H1N1)pdm09, influenza A(H3N2), influenza B, parainfluenza virus (PIV) types 1, 2, and 3, human coronavirus 229E (CoV-229E), human adenovirus (HAdV), herpes simplex virus 1 (HSV 1), and enterovirus (EV). Influenza A viruses grew best in the CACO-2 cell line. Time to observation of CPE was similar for all three cell types but unlike RhMK and MDCK cells, virus-specific morphological changes were indistinguishable in CACO-2 cells. None of the 16 specimens positive for RSV by PCR grew on CACO-2 cells.

CONCLUSIONS

The overall respiratory virus culture isolation rate in CACO-2 cells was significantly higher than that in RhMK or MDCK cells (p < 0.05). CACO-2 cells also supported the growth of some viruses that did not grow in either RhMK or MDCK cells. Except for RSV, CACO-2 cells provide a worthwhile addition to culture algorithms for respiratory specimens.

摘要

背景

原代恒河猴肾细胞（RhMK）可用于检测呼吸道病毒，包括流感病毒和副流感病毒。人结肠腺癌细胞系 CACO-2 之前已用于多种流感病毒的生长，包括季节性、新型和禽流感病毒株。

目的

我们比较了 CACO-2、Madin-Darby 犬肾（MDCK）和 RhMK 细胞在分离表现为流感样疾病（ILI）的患者病毒方面的性能。

研究设计

在基层医疗机构采集ILI 患者的鼻咽标本，在 MDCK、RhMK 和 CACO-2 中进行常规病毒培养。用显微镜检查细胞的细胞病变效应（CPE），并进行免疫荧光检测（IFA）和实时 RT-PCR 进行确认性检测。此外，16 份通过 PCR 检测为呼吸道合胞病毒（RSV）阳性的标本接种在 CACO-2 细胞上。使用 IBM 统计程序进行卡方检验进行统计分析。

结果

在接种的 1031 份呼吸道标本中，331 份（32.1%）在 MDCK 细胞中分离和确认了病毒，304 份（29.5%）在 RhMk 细胞中，433 份（42.0%）在 CACO-2 细胞中分离和确认了病毒。这些病毒包括甲型流感病毒（H1N1）pdm09、甲型流感病毒（H3N2）、乙型流感病毒、副流感病毒（PIV）1、2 和 3 型、人冠状病毒 229E（CoV-229E）、人腺病毒（HAdV）、单纯疱疹病毒 1（HSV 1）和肠道病毒（EV）。甲型流感病毒在 CACO-2 细胞系中生长最好。三种细胞类型观察到 CPE 的时间相似，但与 RhMK 和 MDCK 细胞不同，CACO-2 细胞中病毒特异性形态变化无法区分。通过 PCR 检测为 RSV 阳性的 16 份标本均未在 CACO-2 细胞上生长。