从急性弛缓性麻痹病例中分离1株VP_1编码区Ⅲ/Ⅱ型疫苗重组的脊髓灰质炎病毒  

Identification of a Type 3 / Type 2 Intertypic Vaccine-Related Poliovirus Recombinant Isolated from an Acute Flaccid Paralysis Patient with the Crossover Sites within the VP_1 Capsid Coding Region

作　　者：范明[1] 刘桂艳[1] 周剑惠[1] 王爽[1] 魏雷雷[1] 林琳[1] 周静[2] 田鑫[1] 曹凤瑞[1] 程涛[1] 严冬梅[3] 

机构地区：[1]吉林省疾病预防控制中心,吉林省卫生厅病毒重点实验室,长春130062 [2]白山市疾病预防控制中心,吉林白山134300 [3]中国疾病预防控制中心病毒病预防控制所,卫生部医学病毒学和病毒病重点实验室,世界卫生组织西太平洋区脊髓灰质炎参比实验室,北京102206

出　　处：《中国疫苗和免疫》2015年第2期185-189,共5页Chinese Journal of Vaccines and Immunization

基　　金：吉林省卫生厅急性弛缓性麻痹病例中肠道病毒的研究项目(编号:2008Z086)

摘　　要：目的鉴定吉林省从急性弛缓性麻痹病例粪便标本中分离的1株VP1编码区Ⅲ/Ⅱ型疫苗重组脊髓灰质炎(脊灰)病毒[Vaccine-recombinant Poliovirus(PV),VRPV]。方法粪便标本采自1例2岁吉兰-巴雷综合征病例,使用转人PV受体的小鼠肺细胞系(Mouse Cell Line Expressing the Gene for the Human Cellular Receptor for PV,L20B)细胞和人横纹肌肉瘤(Human Rhabdomyosarcoma,RD)细胞进行病毒分离,阳性标本进行中和试验(Neutralization Test,NT)鉴定血清型,并通过逆转录-聚合酶链反应进行VP1编码区扩增和核苷酸序列测定,利用Bioedit软件对该序列结果与赛宾(Sabin)疫苗株PV进行核苷酸和氨基酸序列比对,分析重组位点的位置以及核苷酸和氨基酸的变异情况。结果该标本接种到RD和L20B细胞后,均发生了肠道病毒特征性病变,经血清学定型,结果为Ⅱ+Ⅲ型PV。VP1编码区核苷酸序列分析表明,其中的Ⅱ型PV与Sabin 2株PV比较,发生了1个核苷酸变异;Ⅲ型PV为VP1编码区Ⅲ型与Ⅱ型VRPV,与相对应的Sabin株PV相比,无核苷酸点变异,在VP1编码区重组的Ⅱ型PV序列为106个核苷酸。血清NT证明,该VRPV仍能被Ⅲ型完全中和,而不能被Ⅱ型中和。结论虽然Ⅲ型PV株中插入了Ⅱ型抗原位点NAg3a,但Ⅲ型PV株总体的型特异性抗原特征未发生改变。PV自然重组很少发生在衣壳蛋白编码区,发现的这株VRPV中和特性与Sabin株PV相比未发生变化。Objective To analyze a type 3 / type 2 intertypic vaccine-related poliovirus（ VRPV） recombinant isolated from acute flaccid paralysis（ AFP） case with the crossover sites within the VP1 capsid coding region in Jilin province. Methods Stool specimens were collected from a 2-year-old patient with Guillain-Barre syndrome,following which virus isolation was performed by inoculating in a mouse cell line expressing the gene for the human cellular receptor for poliovirus（ L20B） and the human rhabdomyosarcoma（ RD） cell line. Positive isolates were identified with the neutralization test and VP1 coding region gene sequence analysis was conducted using reverse transcription-polymerase chain reaction. The BioE dit sequence alignment editor was used to perform sequence alignment of nucleotide and amino acid,and analyze the recombination site location and mutations of nucleotide and amino acid. Results An enterovirus typical cytopathetic effect was observed in both the L20 B and RD cell lines. Type 3 and type 2 mixed poliovirus were identified by the neutralization test. VP1 coding region sequence analysis showed that one nucleotide mutation hap-pened in the type 2 strain,and that the type 3strain was actually a type 3 / type 2 intertypic VRPV recombinant with no point mutation compared with the Sabin 3. Type 2 has 106 nucleotide located in the VP1 coding region. Neutralization testing showed that the recombinant could be only neutralized by type 3 anti-polio serum but not type 2. Conclusion Although type 2 antigen NAg3 a was inserted in type 3 virus VP1 coding region,the recombinant retained Sabin 3-specific characterizations. Natural intertypic capsid recombination in the VP1 capsid coding region is a relatively rare phenomenon; the recombinant we found here still maintained its characteristics of neutralization.

关 键 词：急性弛缓性麻痹 疫苗重组脊髓灰质炎病毒 VP1编码区 

分 类 号：R373.33[医药卫生—病原生物学]