PCR—ELISA在检测血清中丙型肝炎病毒上的应用

建立了一个替代目前使用的套式ＰＣＲ的血清中丙型肝炎病毒ＲＮＡ检测的新方法,该方法只需经过一次ＰＣＲ扩增,即可通过对掺入标记物的ＰＣＲ产物的ＥＬＩＳＡ检测得到与套式ＰＣＲ相同的结果。与套式ＰＣＲ相比,该方法具有耗时少、易操作、较少产生假阳性等特点。     

应用套式PCR选择性卡他热病毒在不同动物体内变异的研究

应用本实验建立的三组套式ＰＣＲ（ＰＣＲ１、２、３）和一组以前报道的套式ＰＣＲ（ＰＣＲ４）,对５９份外周血淋巴细胞（ＰＢＬ）ＤＮＡ样品进行了恶性卡他病毒核酸序列的检测。这些样品来自５１只羊,以及与羊接触而发病的６头牛和２只鹿。除ＰＣＲ４外,其它三组ＰＣＲ都能扩增现有４个角马型ＭＣＦＶ分离株。有６只羊在４组ＰＣＲ中都呈阴性,其余５３份样品经ＰＣＲ４检测均呈阳性。ＰＣＲ１只有从４５只羊体检出ＭＣＦＶＤＮ     

应用循环逆转录PCR技术检测丙型肝炎病毒RNA

循环逆转录（ｃｉｒｃｕｌａｔｏｒｙ ｒｅｖｅｒｓｅ ｔｒａｎｓｃｒｉｐｔｉｏｎ,ＣＲＴ）是线性增长逆转录ｃＤＮＡ产量的一种新技术。为了将该技术用于检测ＨＣＶ ＲＮＡ,通过改变ＣＲＴ的循环次数,结合竞争ＰＣＲ,作出标准曲线。采用１６次ＣＲＴ加３４次循环ＰＣＲ检测了１３６例ＨＣＶ ＥＬＩＳＡ阳性、５４例ＨＣＶ ＥＬＩＳＡ阴性和１０８例临床可疑病人全血标本,并与逆转录ＰＣＲ（ＲＴ－ＰＣＲ）和巢式ＰＣＲ（     

免疫聚合酶链反应技术的建立及其对甲胎蛋白的检测

免疫ＰＣＲ是一种新的具高敏感性的抗原检测技术,它类似传统的ＥＬＩＳＡ法,若与抗体连接的酶用ＤＮＡ片段代替,则该ＤＮＡ片段可用于ＰＣＲ扩增。以链酶亲合素搭桥将生物素标记的抗体与ＤＮＡ相连建立了免疫ＰＣＲ技术,并将其用于检测甲胎蛋白（ＡＦＰ）,其敏感性比ＥＬＩＳＡ法高１０４。因此,免疫ＰＣＲ有可能作为一种具高敏感性的检测手段用于临床早期诊断     

随机引物PCR技术在个体认定及亲权鉴定中的应用

随机引物ＰＣＲ技术（ＡＰ－ＰＣＲ）检测人ＤＮＡ指纹图用于法医学个体认定及亲权鉴定是一个非常有效和经济的方法。应用该技术对１００例无关个体ＡＰ－ＰＣＲ的ＤＮＡ指纹进行分析和１０个肯定亲生关系的家庭进行亲权鉴定,结果该技术的个体识别力非常高,亲权鉴定时的非父排除率也很高,且操作简便、快速,是一个值得推广的方法。     

免疫聚合酶链反应技术的建立及其对甲脂蛋白的检测

免疫ＰＣＲ是一种新的具高敏感性的抗原检测技术,它类似传统的ＥＬＩＳＡ法,若与抗体连接的酶用ＤＮＡ片段代替,则该ＤＮＡ片段可用于ＰＣＲ扩增。因此,免疫ＰＣＲ有可能作为一种具高敏感性的检测手段用于临床早期诊断。     

一步多重PCR同时检测甲型肝炎和脊髓灰质炎病毒研究

建立的一步ＰＣＲ方法即反转录和ＰＣＲ在同一管中进行,同时检测甲型肝炎和脊髓灰质炎病毒病毒ＲＮＡ。实验中对不同的反转录温度以及一步多重ＰＣＲ的特异性和灵敏度进行了探讨。结果表明：４２℃、５０℃反转录时ｐｏｌｉｏ有非特异性条带出现,６０℃反转录特异性较好,而ＨＡＶ在三种不同的反转录温度下均得到牧场划性较好的条带;应用一步ＰＣＲ同时检测两种病毒与检测单一病毒的灵敏度基本一致,但在同等反应条件下后者的反应效率高于前者,特别是在检测ＨＡＶ时。     

应用套式PCR对恶性卡他热病毒在不同动物体内变异的研究

应用本实验建立的三组套式ＰＣＲ（ＰＣＲ１、２、３）和一组以前报道的套式ＰＣＲ（ＰＣＲ４）,对５９份外周血淋巴细胞（ＰＢＬ）ＤＮＡ样品进行了恶性卡他热病毒（Ｍａｌｉｇｎａｎｔｃａｔａｒｒｈａｌｆｅｖｅｒｖｉｒｕｓ,ＭＣＦＶ）核酸序列的检测。这些样品来自５１只羊,以及与羊接触而发病的６头牛和２只鹿。除ＰＣＲ４外,其它三组ＰＣＲ都能扩增现有４个角马型ＭＣＦＶ分离株。有６只羊在４组ＰＣＲ中都呈阴性,其余５３份样品经ＰＣＲ４检测均呈阳性。ＰＣＲ１只能从４５只羊体检出ＭＣＦＶＤＮＡ,未能从牛和鹿体检出病毒ＤＮＡ。ＰＣＲ２检测的所有样品均呈阴性。在ＰＣＲ３扩增中,除２头牛外,其它５１份样品均呈阳性。通过Ｓｏｕｔｈｅｒｎ杂交和限制性酶切分析,对ＰＣＲ１－４产物的特异性进行了鉴定。此外,敏感性实验表明,四组ＰＣＲ的差异也不明显。因此,本实验结果说明ＭＣＦＶ基因组在不同种动物之间发生了变异,羊体内的变异株可能是导致其它反刍动物发病的病原     

应用反转录-套式PCR方法检测肾综合征出血热病人血清中的汉坦病毒特异性RNA

采用５′端生物素标记汉滩病毒特异性寡核苷酸探针,结合磁性分离技术和异硫氰酸胍－酚一步法（胍酚法）提取病毒ＲＮＡ,进行反转录－套式ＰＣＲ,检测肾综合征出血热（ＨＦＲＳ）病人血清样本,胍酚法能检测血清中至少几个ＰＦＵ病毒的ＲＮＡ,比磁珠法敏感１０倍,整个检测过程在５小时完成。应用该方法对１３７份汉滩型和汉城型ＨＦＲＳ病人血清进行检测分型,总阳性率达６０．５０％。其中,病程在７日以内（急性期）的病人血清仍能检测到２２．７３％阳性率。扩增产物经打点杂交检测证实为特异性扩增,并能准确分型。与空斑减少中和试验（ＰＲＮＴ）分型的符合率为１００％。对２０份正常人及２４份非ＨＦＲＳ患者血清进行ＰＣＲ扩增,结果均为阴性。将全部试剂做成试剂盒,为基层单位早期诊断肾综合征出血热病人提供了操作简便、特异、敏感、快速、直接的诊断方法。     

轮状病毒基因重配株血清型鉴定方法的研究

Ａ组轮状病毒中根据基因９的不同目前至少已发现有１３个不同Ｇ血清型,其中能引起人类致病的有Ｇ１－Ｇ４,Ｇ８,Ｇ９和Ｇ１２型。建立可靠的血清型鉴定技术对于轮状病毒疫苗的研制和分子流行病学的研究具有重要意义。本文首次报导了一种鉴定轮状病毒Ｇ血清型的新方法,利用已知有关轮状病毒ＶＰ７基因的序列资料,设计合成了一套鉴定轮状病毒Ｇ血清型的寡核苷酸探针,利用地高辛标记上述探针。待检品经反转录ＰＣＲ扩增后与上述一套寡核苷酸探针分别进行杂交得以确定其血清型。这一方法与目前常用的套式ＰＣＲ方法相比更适合于大量样品的操作而且结果可靠。用这一方法对本实验室组建的四株基因重配疫苗株进行实验,其结果与套式ＰＣＲ方法完全一致。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

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.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

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.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

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.     

肝癌中HBV和HCV基因和抗原的分布及意义

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     

Selection with two alleles and complete dominance

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.