《2006 SARS_associated Coronavirus Replication in Cell Lines》由会员分享，可在线阅读，更多相关《2006 SARS_associated Coronavirus Replication in Cell Lines（6页珍藏版）》请在金锄头文库上搜索。
1、Given the potential for laboratory-associated severe acute respiratory syndromeassociated coronavirus (SARS-CoV) infections, we must know which cell lines are susceptible to the virus. We investigated 21 cell lines rou- tinely used for virus isolation or research. After infection with SARS-CoV, cell
2、s were observed for cytopathic effects, and quantitative real-time polymerase chain reaction was used to measure ongoing viral replication. An indirect immunoflu- orescence assay was also used as a confirmatory test. The study identified 10 new cell lines capable of supporting the replication of SAR
3、S-CoV and confirmed the susceptibility of 4 cell lines previously reported. This study shows that SARS-CoV can be isolated in several cell lines commonly used for diagnostic or research purposes. It also shows that SARS-CoV can achieve high titers in several cell lines, sometimes in the absence of s
4、pecific cytopathic effects. S evere acute respiratory syndrome (SARS) was first observed in 2002 when cases of a life-threatening atyp- ical pneumonia occurred in Guangdong Province, China (1). A novel coronavirus (CoV), designated SARS-CoV, was quickly identified as the etiologic agent (1,2). Altho
5、ugh the origins of the virus have not been estab- lished, evidence suggests that it is an animal virus that was recently transmitted to humans (3). Several wildlife species consumed as delicacies in southern China, includ- ing Himalayan masked palm civets, Chinese ferret badg- ers, and raccoon dogs,
6、 possess antibodies consistent with natural infection with related CoVs (4). Unlike the other currently recognized human CoVs, HCoV-229E, HCoV-OC43, HCoV-NL63, and HKU1, which usually cause mild upper respiratory tract infections and occasionally pneumonia in older adults, neonates, and immunocompro
7、mised patients (58), SARS-CoV causes severe febrile lower respiratory tract illness that leads to pneumonia and acute respiratory distress (9,10). Death from progressive respiratory failure due to alveolar dam- age occurs in 10% of patients with symptomatic infection (2,10). Currently the world is f
8、ree of SARS, but we cannot predict whether the virus will reemerge. The most proba- ble sources of future infections are exposure to animal reservoirs or laboratories where SARS-CoV is manipulat- ed for research purposes. Indeed, since the first epidemic, SARS has occurred on 3 occasions as a result
9、 of breaches in laboratory biosafety procedures (1113). This finding highlights the importance of safely handling SARS-CoV, especially in diagnostic virology laboratories where virus isolation is performed and in research laboratories where infectious virus is handled. SARS-CoV was first isolated in
10、 Vero E6 and FRhK cells injected with clinical specimens as part of early attempts to identify the etiologic agent of SARS (10,14). Simultaneously, these investigations showed that SARS- CoV could not replicate in a number of other cell lines rou- tinely used for respiratory virus isolation. More re
11、cently, additional human and animal cell lines that support SARS- CoV replication have been identified (15). Given the potential for SARS-CoV infection to occur in a laboratory setting, we must be aware of cell lines in which it can repli- cate. Therefore, we investigated the susceptibility of a num
12、ber of cell lines to SARS-CoV. These cells were derived from a variety of species and tissues and included those capable of supporting the replication of respiratory and enteric viruses. Materials and Methods Virus An isolate of SARS-CoV, strain HKU 39849, was pas- saged on 2 occasions in Vero E6 ce
13、lls to establish a high- titer stock that was used in all infectivity experiments. Because SARS-CoV is classified as a risk group level 4 SARSassociated Coronavirus Replication in Cell Lines Matthew Kaye,* Julian Druce,* Thomas Tran,* Renata Kostecki,* Doris Chibo,* Jessica Morris,* Mike Catton,* an
14、d Chris Birch* RESEARCH 128Emerging Infectious Diseases www.cdc.gov/eid Vol. 12, No. 1, January 2006 *Victorian Infectious Diseases Reference Laboratory, North Melbourne, Victoria, Australia pathogen in Australia, all procedures performed with the virus, including infecting cell lines and viral lysi
15、s before RNA extraction, were carried out in a physical contain- ment level 4 (PC4) laboratory. Cell Lines The cell lines investigated for their susceptibility to SARS-CoV are shown in the Table. They were chosen because they were present in our cell repository and were used either routinely or occa
16、sionally for virus isolation attempts as part of diagnostic or research projects. Confluent cells were maintained at 34C in 25-mL flasks (Nunc, Roskilde, Denmark) containing 10 mL appropriate maintenance medium supplemented with fetal bovine serum (FBS) (Thermo Trace, Melbourne, Victoria, Australia), 100 U/mL penicillin, and 100 g/mL strepto- mycin (JRH Biosciences, Lenexa, KS, USA). BGM, FRhK, HEK-293, HEL, Hep G2, L20, MA-104, pCMK, and RD-Acell lines were all maintained in modified Eagle m
