Isolation and molecular characterization of type I and type II feline coronavirus in Malaysia.
Isolation and molecular characterization of type I and type II feline coronavirus in Malaysia.
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DOI：
10.1186/1743-422x-9-278
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发表时间：
2012-11-21
期刊：
影响因子：
4.8
通讯作者：
Tengku Azmi TI
中科院分区：
文献类型：
--
作者：
Amer A;Siti Suri A;Abdul Rahman O;Mohd HB;Faruku B;Saeed S;Tengku Azmi TI
猫传染性腹膜炎病毒（FIPV）和猫肠道冠状病毒（FECV）是世界上两种重要的家猫冠状病毒。虽然FCoV在猫中流行；I型FCoV在细胞培养物上生长的挑剔性质限制了FCoV的组织趋向性和发病机制的进一步研究。虽然有几项研究报告了马来西亚存在口蹄疫病毒的血清学证据，但既没有分离到流行的口蹄疫病毒，也没有确定其生物型。本研究首次报道了从马来西亚自然感染的猫中分离和确定I型和II型FCoV的生物型。在所采集的猫总数中，95%（40/42）为FCoV RT-PCR阳性。将临床样品接种到克兰德尔猫肾细胞（CrFK）和猫catus全胎4细胞（Fcwf-4）中，显示出细胞病变效应（CPE），其特征是合胞细胞形成，随后细胞脱离。利用RT-PCR技术进行FCoV生物型鉴定，本地分离株中ⅰ型和ⅱ型FCoV分别占97.5%和2.5%。这些分离株与来自欧洲、东南亚和美国的FCoV分离株具有高度的序列同源性和系统发育相似性。本研究报道成功分离了局部I型和II型FCoV，在CrFK和Fcwf-4两种类型的细胞培养物中明显形成细胞病变效应，后者的细胞容纳性更强。然而，与在细胞培养中分离病毒相比，RT-PCR在检测可疑样品中的抗原方面更为敏感。目前的研究表明，I型FCoV在马来西亚的猫中更为普遍。
Feline infectious peritonitis virus (FIPV) and feline enteric coronavirus (FECV) are two important coronaviruses of domestic cat worldwide. Although FCoV is prevalent among cats; the fastidious nature of type I FCoV to grow on cell culture has limited further studies on tissue tropism and pathogenesis of FCoV. While several studies reported serological evidence for FCoV in Malaysia, neither the circulating FCoV isolated nor its biotypes determined. This study for the first time, describes the isolation and biotypes determination of type I and type II FCoV from naturally infected cats in Malaysia. Of the total number of cats sampled, 95% (40/42) were RT-PCR positive for FCoV. Inoculation of clinical samples into Crandell feline kidney cells (CrFK), and Feline catus whole fetus-4 cells (Fcwf-4), show cytopathic effect (CPE) characterized by syncytial cells formation and later cell detachment. Differentiation of FCoV biotypes using RT-PCR assay revealed that, 97.5% and 2.5% of local isolates were type I and type II FCoV, respectively. These isolates had high sequence homology and phylogenetic similarity with several FCoV isolates from Europe, South East Asia and USA. This study reported the successful isolation of local type I and type II FCoV evident with formation of cytopathic effects in two types of cell cultures namely the CrFK and Fcwf-4 , where the later cells being more permissive. However, the RT-PCR assay is more sensitive in detecting the antigen in suspected samples as compared to virus isolation in cell culture. The present study indicated that type I FCoV is more prevalent among cats in Malaysia.