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在肾综合征出血热(HFRS)疫苗Vero细胞毒种研制中,将肾综合征出血热(HFRS)病毒PS-6(Ⅰ型)、L99(Ⅱ型)在Vero细胞上连续传代,并对其病毒滴度、免疫原性、传代稳定性等进行检测。结果显示,两株病毒在Vero细胞上传代适应后,病毒滴度达8.5 lgCCID50/m l,免疫原性检查,将病毒灭活制成疫苗免疫家兔的血清中和抗体效价>1:10,其他检测符合规程要求。因此,适应的PS-6(Ⅰ型)和L99(Ⅱ型)毒种可用于制备Vero细胞出血热疫苗。 相似文献
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目的:流感病毒Vero细胞适应株A/Yunnan/1/2005va(H3N2)是一株以Vero细胞为培养基质能高产的病毒株,可用于制备以Vero细胞为基质的流感病毒裂解灭活疫苗;通过在低温下连续传代培养,可选育出流感病毒Vero细胞冷适应株,用于制备Vero细胞流感病毒减毒活疫苗为了便于对Vero细胞冷适应株的进一步研究,建立一个检测该病毒株的ELISA方法.方法:以流感病毒Vero细胞适应株A/Yunnan/1/2005Va (H3N2)的纯化抗原为免疫原制备羊抗A/Yunnan/1/2005Va (H3N2)和鸡抗A/Yunnan/1/2005Va (H3N2)的抗血清.将抗血清先后经硫酸铵沉淀法和Protein G亲和层析柱纯化后,以纯化的羊抗A/Yunnan/1/2005Va(H3N2) IgG为包被抗体,鸡抗A/Yunnan/1/2005Va(H3N2) IgG为第二抗体,测定包被抗体、第二抗体和酶标抗体最佳工作浓度,建立双抗体夹心ELISA检测方法.结果:羊抗A/Yunnan/1/2005Va (H3N2) IgG的最佳工作浓度为5μg/mL,鸡抗A/Yunnan/1/2005Va(H3N2) IgG的最适浓度为10 μg/mL,酶标抗体最佳稀释倍数为1∶4000.对已知的阳性样品,经双抗体夹心ELISA法测定的病毒滴度比血凝方法测定病毒滴度灵敏度高.结论:通过测定包被抗体、第二抗体和酶标抗体最佳工作浓度,建立了双抗体夹心ELISA检测流感病毒Vero细胞适应株病毒含量的方法.该方法操作简单、方便快速、敏感性高,可应用于Vero细胞冷适应株选育时对流感病毒Vero细胞适应株A/Yunnan/1/2005va(H3N2)的检测,对于研制Vero细胞流感减毒活疫苗有重要意义. 相似文献
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不同来源的肾综合征出血热病毒对Vero细胞的致病变作用 总被引：3，自引：0，他引：3
前文报道,肾综合征出血热病毒76-118株能使Vero细胞产生病变。本文报道76-118株和另11株不同来源的肾综合征出血热病毒(H537、A9、H5、R178、HB55、R22、Z10,沟3、L99、A16和J10)对Vero细胞的致病变作用(CPE )。其中除沟3株外,大部分毒株在感染Vero细胞后的第一代即可见明显的CPE。CPE的特点与76-118株相似,主要是感染细胞粘聚、融合,形成网状结构。CPE能被特异性抗HFRS病毒血清和型特异性单克隆抗体所中和抑制,但不能被特异性抗呼肠孤病毒Ⅲ型免疫血清所中和抑制。HFRS病毒对Vero细胞的致病变作用,对进一步研究HFRS病毒的某些生物学特性及实验方法等均有重要意义。 相似文献
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为了解国产浓缩地鼠肾细胞狂犬病疫苗与法国纯化Vero细胞狂犬病疫苗接种人体后中和抗体产生情况。分别用两种疫苗接种 1 0人 ,用小鼠中和试验方法检测中和抗体滴度。国产疫苗五针全程免疫后 30天 (第一针接种后 6 0天 )可 1 0 0 %达到保护水平 ,法国疫苗在第一针接种后 30天 1 0 0 %达到保护水平。第一针接种后 1 4、30、6 0天时 ,前者抗体平均滴度分别是 0 0 6IU /ml、1 .0 2IU/ml、2 .0 7IU/ml;后者抗体平均滴度分别是 0 2 5IU /ml、3.2 4IU/ml、1 1 .86IU/ml,后者比前者产生抗体的滴度高 ,具有显著性差异 (P <0 0 2 )。 相似文献
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对兰州生物制品研究所自行构建的轮状病毒基因重配株G4型LH9株在Vero细胞中传代的稳定性进行研究。将基因重配LH9毒株接种于Vero细胞上从7代传至17代,经电镜检查、病毒滴度测定、病毒基因组RNA电泳图谱分析、基因序列测定分析、轮状病毒型别鉴定及其它相关检定,结果均符合药典要求。证实该毒株为A群G4P〔12〕型轮状病毒,在Vero细胞传代中具有良好的稳定性,为疫苗研制提供安全有效的依据。 相似文献
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在构建了伪狂犬病病毒上海株的缺失载体pgEI-GFP基础上,将pgEI-GFP转染感染了PRV-SH株的BHK-21细胞,待出现80％以上的细胞病变时收获病毒,并以绿色荧光蛋白为标志,通过蚀斑法得到纯化重组病毒gE^-／gI^-／GFP^ 缺失株。研究了该缺失株的的安全性、在细胞上的生长特性以及对断奶仔猪的安全性、免疫原性等生物学特性。试验结果显示,缺失了gE^-和gI^-后,不影响其在RK细胞上的生长状况和病毒的滴度。该疫苗株对小鼠的半数致死量比亲本毒低且对家兔的致死性的时间延长了,这表明该疫苗的毒力比亲本毒有所下降。该缺失株对断奶仔猪安全,无不良接种反应,接种断奶仔猪能抵御高剂量PRV-SH株强毒的感染,攻毒后试验猪的发热期、散毒天数均低于对照组。该缺失株接种仔猪后在试验期间一直维持较高水平的中和抗体。 相似文献
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本文按国际上远交群小鼠的遗传监测和控制的要求,对Ｓｉｂｐ：ＫＭ小鼠进行育种繁殖、遗传背景和生物学特性等多方面的研究。统计分析该小鼠繁殖性能,绘制了生长曲线。测定２４个生化标志基因位点,其中１１个位点呈多态型,平均杂合率为０．２０９。该小鼠下颌骨形态分析的部分判别函数值（占４４％）超过均值±２ＳＤ,呈弥散分布。该小鼠Ｈ－２组织相容合体,具有１０种Ｉ类私有抗原,１７种表现型。此外还测定该小鼠形态数据、部分血液学数据和整体分析,为Ｓｉｂｐ：ＫＭ小鼠推广应用提供技术数据。 相似文献
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为了解国产浓缩地鼠肾细胞狂犬疫苗与法国纯化Ｖｅｒｏ细胞狂犬疫苗接种人体后中和抗体产生情况。分别用两种疫苗接种１０人,用小鼠中和试验方法检测中和抗体滴度。国产疫苗五针全程免疫后３０天（第一针按种后６０天）可１００％达到保护水平,法国疫苗在第一针接种后３０天１００％达到保护水平。第一针接种后１４、３０、６０天时,前者抗体平均滴度分别是０．０６ＩＵ／ｍｌ、１０２ＩＵ／ｍｌ、２．０７ＩＵ／ｍｌ;后者抗体平 相似文献
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Li-chun WANG Song-qing TANG Yan-mei LI Hong-lin ZHAO Cheng-hong DONG Ping-fang CUI Shao-hui MA Yun LIAO Long-ding LIU Qi-han LI 《Virologica Sinica》2010,(2)
The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates s... 相似文献
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目的了解濮阳市手足口病流行病学特征,为制定防控策略提供科学依据。方法通过国家疾病监测信息管理系统收集的全市2008—2012年6月6日手足口病疫情资料进行描述和分析,并对部分病例和重症病例标本进行肠道病毒病原学检测。结果全市共报手足口病16 492例,发病高峰是每年的3-5月(第12～20周),呈典型的单峰型曲线;发病年龄以0～4岁居多;男性多于女性;散居儿童多于托幼机构儿童,爆发病例多发生在托幼机构,手足口病病原有EV71、CoxA16和其他肠道病毒,以EV71和CoxA16为主。结论手足口病发病有明显的季节性、年龄和性别差异,小年龄组儿童是手足口病预防控制重点人群,流行年度和流行季节的优势毒株为EV71,重症患者中EV71占到86.35%;非流行年和季节手足病例主要由CoxA16和其他肠道病毒引起。手足口病防控重点应体现在对病例分类管理上,同时应继续加强重症病例疫情监测和爆发控制。 相似文献
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Chengyuan Wu Guangyan Zhu Fang Qiu Fuli Ren Binbin Lin Dingyu Zhang Qingyu Yang Chaolin Huang 《中国病毒学》2023,38(2):276-284
Enterovirus 71 (EV71) poses a serious threat to human health, with scattered outbreaks worldwide. There are several vaccines against a few EV71 strains but no efficient drug for the treatment of EV71 infection. Therefore, it is urgent and of significance to develop anti-EV71 drugs. Here, we found that PLX8394, a RAF inhibitor, possesses high antiviral activity against EV71 in vitro, being superior to the traditional clinical drug ribavirin. Moreover, PLX8394 exhibits broad-spectrum antiviral activity against enteroviruses. Notably, in a suckling mouse model, PLX8394 provided a 70% protection rate for EV71-infected mice, reduced the viral load in liver and heart tissues, and relieved the inflammatory response. A mechanistic study showed that PLX8394 inhibited EV71 by suppressing the RAF/MEK/ERK signaling pathway. Thus, PLX8394 lays a foundation for the development of new drugs against EV71. 相似文献
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Qiu-Yan Zhang Jia-Qi Li Qi Li Yang Zhang Zhe-Rui Zhang Xiao-Dan Li Hong-Qing Zhang Cheng-Lin Deng Feng-Xia Yang Yi Xu Bo Zhang 《中国病毒学》2024,39(2):301-308
Hand, foot, and mouth disease (HFMD) is a common pediatric illness mainly caused by enteroviruses, which are important human pathogens. Currently, there are no available antiviral agents for the therapy of enterovirus infection. In this study, an excellent high-content antiviral screening system utilizing the EV-A71-eGFP reporter virus was developed. Using this screening system, we screened a drug library containing 1042 natural compounds to identify potential EV-A71 inhibitors. Fangchinoline (FAN), a bis-benzylisoquinoline alkaloid, exhibits potential inhibitory effects against various enteroviruses that cause HFMD, such as EV-A71, CV-A10, CV-B3 and CV-A16. Further investigations revealed that FAN targets the early stage of the enterovirus life cycle. Through the selection of FAN-resistant EV-A71 viruses, we demonstrated that the VP1 protein could be a potential target of FAN, as two mutations in VP1 (E145G and V258I) resulted in viral resistance to FAN. Our research suggests that FAN is an efficient inhibitor of EV-A71 and has the potential to be a broad-spectrum antiviral drug against human enteroviruses. 相似文献
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Shao-hui Ma Jian-sheng Liu Jing-jing Wang Hai-jing Shi Hui-juan Yang Jun-ying Chen Long-ding Liu Qi-han Li 《中国病毒学》2009,24(3):162-170
Enterovirus 71 (EV71) is a common cause of Hand, foot, and mouth disease (HFMD) and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences (891 nucleotides) from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous to members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine(H) at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains. 相似文献
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Yonghong Zhou Chongchen Zhou Kai Wang Qi Qiu Yibing Cheng Yu Li Peng Cui Lu Liang Peng Li Xiaowei Deng Lili Wang Wen Zheng Hui Gong Fang Wang Meng Xu Justin Jang Hann Chu Lance Turtle Hongjie Yu 《中国病毒学》2023,38(2):268-275
Hand, foot and mouth disease (HFMD) is a major public health problem among children in the Asia-Pacific region. The optimal specimen for HFMD virological diagnosis remains unclear. Enterovirus A71 (EV-A71) neutralizing antibody titres detected in paired sera were considered the reference standard for calculating the sensitivity, specificity, positive and negative predictive value of throat swabs, rectal swabs, stool, blood samples and cerebrospinal fluid (CSF) by RT-PCR or ELISA assay. In this study, clinical samples from 276 HFMD patients were collected for analysing the sensitivity of different kind of specimens. Our results showed that stool had the highest sensitivity (88%, 95% CI: 74%–96%) and agreement with the reference standard (91%). The order of diagnostic yield for EV-A71 infection was stool sample ≥ rectal swab > throat swab > blood sample > CSF sample, and using a combination of clinical samples improved sensitivity for enterovirus detection. The sensitivity of ELISA for IgM antibody detection in sterile-site specimens was significantly higher than that of RT-PCR (serum/plasma: 62% vs. 2%, CSF: 47% vs. 0%) (P < 0.002). In conclusion, our results suggest that stool has the highest diagnostic yield for EV-A71-infected HFMD. If stool is unavailable, rectal swabs can be collected to achieve a similar diagnostic yield. Otherwise, throat swabs may be useful in detecting positive samples. Although IgM in blood or CSF is diagnostically accurate, it lacks sensitivity, missing 40%–50% of cases. The higher proportion of severe cases and shorter interval between onset and sampling contributed to the increase in congruency between clinical testing and the serological reference standard. 相似文献
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Rong-Rong Zhang Meng-Jiao He Chao Zhou Yan-Peng Xu Wei Tang Tian-Shu Cao Zheng-Jian Wang Mei Wu Tao Ming Yi-Jiao Huang Meng-Xu Sun Hui Zhao Yong-Qiang Deng Xiao-Feng Li Bin Wang Qing Ye Cheng-Feng Qin 《中国病毒学》2024,39(5):812-820
Human Enterovirus 71 (EV71) has emerged as one of the predominant causative agents of hand, foot and mouth disease (HFMD) with global impact. Despite the inactivated vaccine being licensed, other vaccine candidates based on advanced technology platforms are under development. In this report, we rationally designed and constructed two DNA-launched live attenuated vaccine candidates (pDL-EV71) under the control of specific promoters. In vitro and in vivo transfection with pDL-EV71 driven by the CMV promoter successfully yielded fully infectious EV71. More importantly, the administration of pDL-EV71 did not cause clinical symptoms following intracranial or intramuscular inoculation in neonatal and IFNα/βR-/- mice, demonstrating its safety profile. Moreover, a single-dose or two-dose immunization with pDL-EV71 elicited robust neutralizing antibodies against EV71 as well as an antigen-specific cellular response in mice. A single-dose immunization with 10 μg of pDL-EV71 conferred complete protection against lethal EV71 infection in neonates born to immunized maternal mice. Overall, our present results demonstrate that pDL-EV71 is a safe and effective vaccine candidate against EV71 for further development. 相似文献
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人免疫球蛋白中肠道病毒71型中和抗体效价的测定 总被引：5，自引：0，他引：5
采用经典微量细胞病变法,应用近两年分离自中国手足口病(HFMD)高发区的3株肠道病毒71型(EV71)病毒株,对中国不同血液制品厂家生产的35批人免疫球蛋白制品进行抗-EV71中和效价检测。结果显示,3株不同EV71病毒株间的抗-EV71中和效价差异均在4倍以内,差异无显著的统计学意义(F=2.323,P0.05)。根据这3株毒株检测结果判定,35批人免疫球蛋白的抗-EV71均为阳性,肌肉注射用免疫球蛋白(简称肌丙)的抗-EV71-GMTs(525.9)显著高于静脉注射用免疫球蛋白(简称静丙)的GMTs(252.3,F=66.518,P0.01)。30批静丙的抗-EV71-GMTs中和效价分布在128.0~384.0之间。应开展原料血浆中抗-EV71中和效价的筛选,研制高效价的EV71特异性免疫球蛋白制品,用于HFMD的治疗和预防。 相似文献
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Zhang Qiuhan Li Siliang Lei Ping Li Zixian Chen Feifei Chen Qi Wang Yulu Gong Jiami Tang Qi Liu Xinjin Lan Ke Wu Shuwen 《中国病毒学》2021,36(6):1387-1399
Virologica Sinica - Similar to that of other enteroviruses, the replication of enterovirus 71 (EV71) occurs on rearranged membranous structures called replication organelles (ROs).... 相似文献