关键词： Animals   Base Sequence   Cell Line   Cercopithecus aethiops   DNA Mutational Analysis   DNA, Viral   Evolution   Genetic Variation   Introns   Macaca mulatta   Molecular Sequence Data   Mutation   Poliomyelitis/microbiology   Poliomyelitis/microbiology   Poliomyelitis/prevention & control   Poliomyelitis/prevention & control   Poliovirus Vaccine, Oral/genetics   Polymerase Chain Reaction   Vero Cells   Virulence  

摘要： Mutants consistently accumulating in Sabin 1 poliovirus during serial passaging in vitro were identified by sequence heterogeneity assay and quantitated using mutant analysis by PCR and restriction enzyme cleavage (MAPREC). Only four unstable genomic sites were identified in virus passaged 10 times in African green monkey kidney (AGMK) cells, and eight sites in virus passaged in Vero cells. Mutations accumulated both in untranslated regions of RNA (nucleotides 480, 525 and 7441) and in coding sequences, as missense (nucleotides 1449, 4944, and 6203) or silent (nucleotides 1123 and 1141) mutations. The most prominent selectable mutations were found at complementary nucleotides 480 and 525 of the 5'-untranslated region (5'-UTR) of the Sabin strain, changing the G:U pair in F-domain to either A:U or G:C variants. These two variants have been shown previously to have an increased neurovirulence in monkeys. The G:C variant accumulated during passage in Vero cells, while A:U variant accumulated in CV-1 cells. Virus passaged in AGMK cells accumulated both variants. Higher temperature (37 instead of 34 degrees) strongly favored selection of mutants in Vero cells, had a smaller effect on mutant accumulation in AGMK cells, and had no effect in CV-1 cells. Monopools of type 1 oral poliovirus vaccine (OPV) made by seven manufacturers were found to contain both 480-A and 525-C revertants at a combined level of 1.1-2.7%. Viral samples with increased amounts of these revertants had higher neurovirulence in monkeys. Our results suggest that quantitation of these reversions by MAPREC may be prognostic for results of the monkey neurovirulence test (MNVT) and can be used for monitoring type 1 OPV consistency. (C) 1994 Academic Press, Inc.

关键词： 社会保障水平   保障形式   乡镇卫生院   儿童计划免疫   百白破三联制剂   劳保医疗   精神文明建设   预防保健   脊髓灰质炎疫苗   妇幼保健机构  

关键词： 脊髓灰质炎   减毒活疫苗   病毒   序列  

摘要： 对中国脊髓灰质炎Ⅱ型减毒活疫苗株──中Ⅱ１７和Ⅲ型减毒活疫苗株──中Ⅲ２的ＶＰ１段部份核酸片段进行序列分析，并分别与相应的Ｓａｂｉｎ２和Ｓａｂｉｎ３株有关片段的核苷酸序列进行比较。在４１６个核苷酸序列中，中Ⅱ１７与Ｓａｂｉｎ２有９９．３％同源性，中Ⅲ２与Ｓａｂｉｎ３株完全一致。从我国服用国产活疫苗地区急性弛缓性麻痹病例中分离到的部份２型疫苗株，分别与中Ⅱ１７和Ｓａｂｉｎ２相比较，发现与中Ⅱ１７株更为近缘，海南、山东、湖南株分别只有０、１或２个核苷酸的改变。

关键词： 脊髓灰质病毒   蚀斑法   微滴法   滴定   疫苗  

摘要： 用蚀斑法和微滴法滴定脊髓灰质炎病毒的比较戴永祥，李克坚，王明秀，姜述德（中国医学科学院医学生物学研究所，昆明６５０１０７）口服脊髓灰质炎活疫苗（ＯＰＶ）生产，一直使用原代猴肾细胞，由于猴子来源困难，而且污染猴病毒，直接影响到疫苗生产。用人二倍体细胞代...

关键词： 原发性免疫缺陷病   红细胞免疫功能   脊髓灰质炎病毒   红细胞C3b受体   流行性脑脊髓膜炎   风疹病毒   麻疹病毒   免疫球蛋自   临床儿科杂志   花环试验  

关键词： 脊髓灰质炎病毒   胆汁   胆道疾病  

摘要： 从胆道疾患病人采集的胆汁标本，用组织培养方法进行病毒分离。从６例胆道疾患病人的胆汁中分离出脊髓灰质炎病毒，经用标准免疫血清中和试验，证实为Ⅱ型脊髓灰质炎病毒。表明脊髓灰质炎病毒可以在胆道系统中存在，并可能是胆道感染的原因之一。对胆汁对组织培养细胞的毒性作用及其去除方法作了探讨。

摘要： Although mass vaccination programs have resulted in the eradication of a number of human infectious diseases, vaccine contamination has been a persistent concern. In particular, it is now known that the early polio vaccines were contaminated with at least one monkey virus, SV40. The transfer of monkey viruses to man via contaminated vaccines is particularly relevant to the acquired immunodeficiency syndrome (AIDS), since the causative agent of AIDS, human immunodeficiency virus (HIV), is thought to be derived from a simian precursor virus. Furthermore, human infection with this virus appears to be a relatively recent event. We hypothesize that the AIDS pandemic may have originated with a contaminated polio vaccine that was administered to inhabitants of Equatorial Africa from 1957 to 1959. The mechanism of evolution of HIV from this vaccine remains to be determined.

关键词： Animals   Cell Line   Cell Membrane/metabolism   Humans   Kinetics   Membrane Proteins   Poliovirus/metabolism   Precipitin Tests   Precipitin Tests   Receptors, Virus/analysis   Receptors, Virus/metabolism   Species Specificity   Tumor Cells, Cultured  

摘要： The interaction of poliovirus with its cellular binding sites was characterized by using a receptor-excess silicon oil partition assay. Poliovirus type 1 Mahoney [PV1(M)] binding to HeLa cells fits a theoretical simple bimolecular noncooperative binding curve with an equilibrium dissociation constant (K-d) of 4.3 X 10(7) ***(-1) at 4 degrees, or 2.1 x 10(-10) M, assuming 3000 virus binding sites/cell. The association rate of complex formation was measured to be 3.6 x 10(-9) ***(-1).min(-1) (7.2 X 108 M(-1).min(-1)) and the dissociation rate calculated to be 1.5 x 10(-1) min(-1), giving the complex a half-life of 4.5 min. The equilibrium dissociation constant, association rate, and dissociation rate were also measured for the binding of the attenuated poliovirus type 3 Sabin strain [PV3(S)] to HeLa cells. PV3(s) bound HeLa cells with a K-d of 3.3 X 10(7) ***(-1) (1.6 x 10(-10) M), an association rate of 4.1 x 10(-9) ***(-1).min(-1) (8.2 x 10(8) M(-1).min(-1)), and a dissociation rate calculated to be 1.4 X 10(-1) min(-1), giving the complex a half-life of 5.1 min. Thus the virulent and avirulent strains of poliovirus bind HeLa cells with nearly identical binding constants and rate constants. Equilibrium binding constants for PV1(M) to various other cell types varied from a high affinity of 4.1 x 10(6) ***(-1) for JA-1 cells to a low affinity of 7 x 10(7) ***(-1) for NGP cells. Comparison of the amount of immunoprecipitated poliovirus receptor suggests that the weak virus binding of NGP cells may be due to a limited number of receptors expressed on the cell surface of these cells, (C) 1994 Academic Press, Inc.

关键词： 脊髓灰质炎   全程免疫   病因   儿童  

摘要： 为探讨脊髓灰质炎（简称脊灰）全程免疫病例发病的危险因素，我们于１９９２年６～１０月份在广西、河南、江苏三省的部分县、乡中进行了一次１：２配比的病例对照研究，通过对收集到的资料进行单因素和多因素条件Ｌｏｇｉｓｔｉｃ回归分析认为，引起全民免疫儿童发病的危险因素包括团体因素和个体因素两大类。团体因素主要指与冷链及卫生服务有关的因素，为脊灰全程服苗病例发病所特有的危险因素。个体因素主要指一些社会行为因素，为免疫及未免疫儿童共同的发病危险因素。此外，服用５次及５次以上脊灰疫苗是一个保护性因素，提示在不断提高接种率的同时，应注意提高接种质量。

关键词： POLIOVIRUS   SITE-SPECIFIC POLYMERASE CHAIN REACTION   POLIO VACCINE   POINT MUTATIONS   ATTENUATION   NEUROVIRULENCE  

摘要： One of the problems raised by the use of the attenuated oral poliovirus vaccine (OPV) Sabin strains is the generic instability of the attenuated phenotype upon multiplication in vivo. Nucleotide sites critical for attenuation were identified for each of the three poliovirus serotypes. One important position lies in the 5' non-coding region of the genome of each of the three OPV strains, at nucleotide 480 in type 1, 481 in type 2 and 472 in type 3. Point mutations at these positions were usually selected upon multiplication in vivo as substitutions of the vaccine-type residue. The reversion was found to correlate with an increased degree of neurovirulence. To screen easily for this mutation in a great number of strains, we developed a site-specific polymerase chain reaction method based on the property of the Tag polymerase to elongate only primers with a perfect homology at the 3' extremity. We screened for this mutation in Jive type I and nine type 2 polio vaccine-derived strains isolated from vaccine-associated paralytic poliomyelitis (VAPP) cases and in 16 such strains isolated from healthy vaccinees. All 14 strains isolated from VAPP presented the reversion. Of the eight pairs of type I isolates from healthy vaccinees, four presented the reversion 3 days after vaccine administration and all but one at 7 days postvaccination. These results support the involvement of the 5' non-coding specific nucleotide sites in the reversion to neurovirulence of attenuated polio vaccine strains upon multiplication in the human gut. The mutation is rapidly selected in type I vaccine poliovirus, which is reputed to be the most stable of the three polio vaccine strains.