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1.
中国脊髓灰质炎病毒疫苗株基因变异的分析   总被引:12,自引:0,他引:12  
李杰  张礼璧 《病毒学报》1996,12(4):291-298
从急性弛缓性麻痹(AFP)病例分离的32株脊髓灰质炎(脊灰)病毒,在VP1编码区,经PCR-RFLP方法鉴定,并经核酸测序进一步证实力疫苗株,其中5株为I型疫苗,16株Ⅱ型疫苗株,11株Ⅲ型疫苗株,以同样的分析方法检测这些毒株基因组的3D聚合酶编码区,发现2株I型疫苗株在3D区的431个核苷酸序列为野毒序列,即这2株Sabin1基因型(VP1)与野毒基因型(3D)重组株;其余3株I型疫苗株未发现基  相似文献   

2.
PCR法检测外环境水中脊髓灰质炎病毒I型基因的研究   总被引:1,自引:0,他引:1  
应用聚合酶链反应(PCR)技术检测外环境水中分离的脊髓灰质炎病毒I型(PVI)毒株基因型,发现所分离的22株病毒,均为疫苗SabinI相关株,脊髓灰质炎病毒的温度敏感(T特征)试验亦提示为弱毒标,与PCR试验相吻合,表明实施强化免疫和常规免疫后,外环境中的脊髓灰质炎病毒已被疫苗相关株循环所取代,同时证实用了PCR作为相环境中PVI病毒株的基因型分析的检测方法是特异和敏感的。  相似文献   

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以来自哈尔滨传染性法氏囊病病毒(IBDV) 强毒株(Harbin 毒株,H) 的基因组RNA为模板,用反转录聚合酶链反应(RT- PCR) 的方法得到了其A 节段的全长cDNA 片段,分5'端(1 659bp) 和3'端(1 444bp) 上下两段分别克隆到pGEMB○R - T 载体上,测定了其核苷酸顺序,在长为3 101 bp 中含有两个阅读框ORFA1 和ORFA2 ,分别编码1 012 个氨基酸的前体蛋白(VP2 - 4 -3) 和145 个氨基酸的VP5,ORFA1 和ORFA2 有部分的重叠。将核苷酸序列及推测出的氨基酸序列与已报道的IBDV 血清Ⅰ型和Ⅱ型毒株的相应序列进行了比较,结果表明:H 毒株与其它血清Ⅰ型毒株之间,在核苷酸水平上存在25bp - 267bp 的差异;在氨基酸水平上存在17 ~40 个氨基酸的差异。在VP2 - 4 - 3 内比较显示,H 毒株与P2 、Cu- 1 之间氨基酸的差异最小为1 .7% ,H 毒株与UK661 之间氨基酸的差异最大为3 .9 % 。变异主要发生在VP2 的可变区(206 - 350 位氨基酸) ,在H 毒株所特有的12 个氨基酸当中,该区就占5 个,代表1 .76 % 的变异。VP4、VP3 和VP5区各有  相似文献   

4.
中国脊髓灰质炎Ⅰ型野毒株的PCR-RFLP法分析   总被引:1,自引:1,他引:0  
近年来我国流行地区分离的脊髓灰质炎病毒多为Ⅰ型野毒。本文报导对我国一些省、自治区防疫站从急性驰缓性麻痹病人粪便中分离的77株Ⅰ型野毒株,经PCR-RFLP法分析结果,发现不同地区分离的野毒株在电泳图像上有明显差异。根据电泳图像所显示的三种酶切条带的数目,可分为14个类型,而且同一省、自治区内不同地区(如新疆),或同一地区不同年份(如广东)的毒株也存在着明显差异。但在某些相邻省份如广东省与海南省,92年流行毒株电泳图像完全一致。说明本法虽不如核酸序列分析精确,但在一定程度上也能反映毒株间的差异,有助于说明流行株的亲疏关系,可用于研究毒株的分子流行病学。  相似文献   

5.
及时发现脊髓灰质炎(脊灰)野病毒,是消灭脊灰工作中病毒学监测的首要任务。近期我国存在4个脊灰Ⅰ型野病毒基因型,其中P1/CHN-JX89和P1/CHN-R91为两个主要的流行基因型。在分析了我国大量脊灰野病毒VP1核酸序列的基础上,选取了1338JX89病毒VP15′端的96个核苷酸片段(2480-2575),经PCR扩增,克隆至pUC/T7质粒,线性化后,用T7RNA多聚酶制备RNA探针,并将Dig标记物渗入探针分子中。经杂交试验,两个主要基因型病毒全部与此探针呈阳性反应,而其它基因型和疫苗相关病毒均为阴性反应,体现了较好的特异性与敏感性,而且实验结果由核酸序列分析等证实。脊灰野毒探针的应用在国际上尚未见报道,它克服了疫苗探针杂交中容易遗漏野毒株的缺点,直接查出野病毒。这对疫苗株中混有少量野毒株的分离物有重要意义。  相似文献   

6.
从我国内蒙古地区流行的犬细小病毒病病犬的肠溶物中分离提纯犬细小病毒(CPV)。提取病毒基因组DNA,并以此DNA为模板,采用人工合成的引物进行PCR扩增,PCR产物经BamHI、SacI双酶切后,克隆于pUC19质粒的BamHI/SacI位点。重组质粒pUCVP2经PCR鉴定、限制酶切分析和序列分析,结果表明:获得了犬细小病毒内蒙株(CPV-IM)VP2基因的全长克隆,VP2基因全长1755nt,  相似文献   

7.
陈杰  苏玲中 《病毒学报》1998,14(3):240-245
使用PCR技术对14份广西HIV-1阳性感染者外周血单核细胞(PBMCs)样品进行扩增,获得HIV-1膜蛋白(env)基因的核酸片段,并对其C2-V3及邻区350 ̄450个核苷酸序列进行了测定和分析。结果表明,14份样品中9份为泰国B(B')亚型,5份为E亚型毒株。其中B'亚型毒株的基因离散率为4.2%,与A-E参考亚型及部分B亚型代表株序列相比较,与包括泰国、缅甸及云南德宏在内的B亚型毒株序列十  相似文献   

8.
用RT-PCR和RFLP对禽传染性支气管炎病毒中国分离株的分型   总被引:7,自引:0,他引:7  
用RT-PCR方法获取了禽传染性支气管炎病毒(IBV)标准毒株M41、H52、A5968和国内分离株D41、F、G的S1基因,对它们做RFLP分析,发现D41、F株属于马萨诸塞血清型、G株为变异株。对用RT-PCR和RFLP来区分IBV的分型方法的应用理论和前景进行了论述。  相似文献   

9.
多重PCR法检测对皮下和造血器官坏死杆状病毒   总被引:1,自引:0,他引:1  
夏春  黄捷 《病毒学报》2000,16(3):262-265
根据对虾皮下和造血坏死杆状病毒(HHNBV)HHNBV-XIA靶基因序列设计了4个多重PCR法用引物(P1、P2、P3、P4)。采用了五种引物组合形式分别对HHNBV-XIA、HHNBV基因和PRDV-JAPAN片段进行了物异扩增,建立了多重PCR检测HHNBV方法。通过优化多重PCR法检测HHNBV条件,可从阳性感染的中国对虾fg级DNA中定性检测出皮下和造血器官坏死杆状病毒。  相似文献   

10.
传染性法氏囊病病毒CJ-801bkf毒株VP2 cDNA基因结构的分析   总被引:2,自引:0,他引:2  
以传染性法氏囊病病毒(IBDN)中国毒株CJ-801bkf的基因组A片段dsRMA为模板,经反向转录和PCR扩增,克隆了保护性抗原VP2cDNA基因。经Sanger法测序,确定了VP2cDNA基因有1484个核苷酸,推测了VP2氨基酸的顺序,与已报导的6株IBDV毒株CuI、PBG98、52/70、002-73、STC和VariantE的VP2区做了比较,证明:克隆的CJ-801bkfVP2cDNA基因是全长的,含有正确的起始密码子ATG;CJ-801bkf与毒株Cul和pBG98同源性最高;CJ-801bkfVP2高可变区内两个亲水区的氨基酸顺序与CuI、PBG98、52/70、002-73和STC完全相同;7肽区内第三个丝氨酸残基则变异为精氨酸。这些结果提示,中国CJ-801bkf毒株应属标准血清I型IBDV弱毒株。  相似文献   

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It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.  相似文献   

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Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.  相似文献   

18.
Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.  相似文献   

19.
肝癌中HBV和HCV基因和抗原的分布及意义   总被引:1,自引:0,他引:1  
采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细  相似文献   

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For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.  相似文献   

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