核酸电泳法检测轮状病毒(摘要)

轮状病毒(Rota Virus)引起的腹泻,粪便中含有大量轮状病毒(可达10~9—10~(11)/ml),可从粪便中直接提取RNA,用垂直板聚丙烯酰胺淀胶电泳法检测轮状病毒 RNA 基因组(简称核酸电泳法)、根据特有的电泳图型(显11条带)进行诊断。1984年我站在研究病毒性腹泻时首次建立起此方法。采用电镜观察轮状病毒阳性与阴性粪便各20份,其核酸电泳检测结果:电镜阳性者均见到清晰的11条带,即为核酸电泳阳性;电镜阴性者均未见到显带,为核酸电泳阴性。显     

人的轮状病毒核酸的提取及鉴定

轮状病毒是婴儿及幼畜腹泻的主要病原之一。轮状病毒核酸是具有11个片段的 RNA 分子。我们从患儿粪便中提取轮状病毒核酸,利用聚丙烯酰胺凝胶电泳法分离 RNA 片段及分子量测定。用分光光度法测定了核酸的紫外吸收,并对核酸及病毒颗粒进行了电镜观察。     

引起婴幼儿腹泻的成人腹泻轮状病毒

从婴幼儿急性腹泻患者粪便中发现6株新轮状病毒,ELISA和免疫电镜证明属B组成人腹泻轮状病毒(ADRV),但其基因组核酸电泳图型为4:2:3:2,与典型ADRV(4:2:2:3)不同,而与婴幼儿腹泻A组轮状病毒相似,可能是ADRV基因组变异株。     

四川地区小儿轮状病毒腹泻病原流行分析

目的：为了解小儿轮状病毒腹泻病原流行情况，方法：采集四川地区９４９例３岁以下疑诊为冬季腹泻患儿全大便，用ＥＬＩＳＡ法，直接电镜和核酸电泳进行轮的和形态的检测。结果：轮状病毒捩的阳性７１．１％；直接电镜阳性为８０．０％；核酸电泳阳性为８７．５％。结论：小儿轮状病毒感洒季节为每年１１月一次年２月，流行高峰为１２月。１岁以下患儿占７５．６％，性别、男女比例为２：１。     

ELISA检测盒对小儿腹泻粪便中轮状病毒的检测

轮状病毒是一种严重影响儿童健康的急性腹泻病原体,常规要用电镜或免疫电镜检查,给诊断带来不便和困难,ELISA法则较为简便。1984～1985年间,我们采用双盲法对小儿腹泻患者粪便用ELISA法及直接电镜检查法(EM)进行了轮状病毒的检测,发现两种办法的检查结果相似,现报道如下:一、标本:系儿科门诊急性腹泻患者     

轮状病毒肠炎病原的快速诊断

轮状病毒(RV)是秋冬季婴幼儿腹泻的主要病原。目前可用电镜、ELISA、核酸电泳、乳胶凝集等技术从患儿粪便中检出病毒。其中以ELISA法应用最广、也较敏感,且试剂盒易得。我们采用ELISA(即A组轮状病毒单克隆抗体快速一步法)检测粪便RV抗原。该方法是一种诊断轮状病毒肠炎病原的快速、简便方法。现将结果报告如下:     

小儿消化功能紊乱性疾病(3)

(接上期) 10.什么是轮状病毒腹泻 轮状病毒是引起婴幼儿腹泻的主要病原之一,由于腹泻多发生于秋冬寒冷季节,过去又称为秋季腹泻.1973年应用电镜检查患儿的粪便发现一种特殊车轮状外形的病毒,命名为轮状病毒.     

人轮状病毒的基因分析和血清型鉴定

本文报道1988年6月～1985年5月期间经电镜或/和ELISA证实含有轮状病毒的28份上海婴幼儿腹泻粪便标本、2份澳大利亚腹泻患儿的粪便标本和1份外地成人腹泻粪便标本,提取核酸进行聚丙烯酰胺凝胶电泳,比较它们的电泳图形。上海28份标本中19份呈长型,3份呈短型,长型有4种电泳图形(L_1～k),短型有2种(S_1,S_2),L_1占多数(10/22),其特征为第7、8、9条三条分开,但7和8之间间距小于8和9之间间距。澳大利亚2份标本的图形与上海的L_1相似。一份外地成人标本电泳条带亦为11条,但图形和其他电泳图形不同,而与国内1982年后发现的成人轮状病毒相同。有4株轮状病毒经用型特异性的单克隆抗体鉴定其血清型为1型,他们的电泳图形属长型。     

核酸电泳分析延吉市爆发流行的轮状病毒基因片段

本文报道核酸电泳分析1986年5月延吉市爆发流行的成人腹泻轮状病毒基因片段。我们收集了22份成人腹泻患者的粪便,在电镜下从11份样品中检出轮状病毒颗料。用聚丙烯酰胺凝胶电泳法13例获得具有特征的11片段 RNA 基因组电泳图谱。这种 RNA 基因片段不同于从婴幼儿腹泻患者中分离出来的轮状病毒 RNA 基因片段的电泳图谱,却同最近洪涛等报导的成人腹泻轮状病毒相符。     

成人流行性腹泻病原基因组的电泳分析

通过基因组电泳分析发现成人流行性腹泻病因是由11个分节段的双链RNA组成,其电泳带的分布模式与典型的以及报道过的轮状病毒皆有明显区别,电镜下呈典型轮状病毒形态,而血清学上无同属的共同抗原,故可确认它是一种新的轮状病毒。     

Rotavirus RNA electropherotype in different states in Malaysia for the year 2000 and 2001

A total of 157 stool samples were examined for Group A rotaviruses in diarrheic children admitted to 8 different major hospitals in Malaysia. The overall incidence rate in this study was 19.7% (31 of 157) with a variation of 9.5% to 39.1% in different locations. Majority of the infections detected were in those under 2 years of age and there were fewer admissions in the older age group. The stool samples were initially screened for rotavirus Group A by latex agglutination method and followed by RNA electrophoresis. The size and the characteristics wheel-shaped morphology of the viral preparations when examined by electron-microscopy further confirmed the presence of rotaviruses in the positive stool samples. Analysis of the RNA pattern showed that majority of the isolates, 51.6% (16 of 31) were Type IIC ('long' with comigration of RNA segments 7 and 8), 35.5% (11 of 31) with Type IIG ('long' with comigration of segments 7, 8, 9), 9.7% (3 of 31) with Type IG ('short' with comigration of RNA segments 7, 8, 9) and 3.2% (1 of 31) of mixed or atypical pattern. It appeared that over a 12 year interval, only one new or unusual rotavirus electropherotype was found. This is the first comprehensive report on the electropherotypes of rotaviruses covering eight different geographical locations in Malaysia and the data obtained is useful for understanding the geographic distribution and types of rotaviruses transmitting in Malaysia.     

用PAGE电泳法检测腹泻婴幼儿粪便中轮状病毒RNA

目的　探讨婴幼儿轮状病毒感染诊断方法。方法　用聚丙烯酰胺凝胶电泳法(PAGE)检测轮状病毒RNA。结果　 41份腹泻婴幼儿粪便标本中有 16份为RNA阳性 ,阳性率为 39%。结论　用PAGE电泳方法检测轮状病毒RNA准确可靠 ,不易出现假阳性     

轮状病毒培养纯化及重组子代分析

人轮状病毒Wa和Hu7株细胞培养条件中,胰蛋白酶处理和旋转培养,在病毒增殖和细胞病变生成过程中起关键作用。通过对一株人轮状病毒FH4232株与猴轮状病毒SA11株的混合感染株的体外培养和空斑纯化,发现子代克隆中有2株FH4232与SA11株生成的重组子代。重组子代病毒株的第5、11条泳带来自亲代FH42321株,其余多条则来自亲代SA11株。体外轮状病毒重组子代的发现对研究其基因重组变异有重要意义,而且为疫苗制备和应用提供有效途径。     

成人轮状病毒腹泻

采用聚丙烯酰胺凝胶电泳及硝酸银染色法,检测近一年来我院162例成人非细菌性肠炎的水样大便,发现19例样本是普通轮状病毒RNA区带;另1例呈“成人腹泻轮状病毒”区带。讨论了其临床表现和一年内的分布情况。     

Virus infection and hand papular dermatoses in young children.

Four strains of enterovirus and 1 strain of adenovirus were isolated from stool and pharyngeal swabs by inoculation into primary human embryonic kidney cell culture in 25 cases of dorsum manus infantile papular dermatitis. Two strains of Coxsackie A-9 virus and 1 strain of Echovirus-25 were identified. With the isolated virus as antigen, a matched pair study by sera antibody test was carried out in 43 children with papular dermatitis and 40 normal individuals as controls. In 26 cases of Coxsackie A-9 virus positive papular dermatitis, a 4-fold rise (> 1:1.6) in the relative serum antibody was observed in 80.77%, while no high levels were observed in patients with Echovirus-25 and adenovirus dermatitis (< 1:16). The authors can reasonably conclude that epidemic Coxsackie A-9 virus infection is obviously related to infantile papular dermatitis and is probably the main cause of the disease.

     

FURTHER STUDIES ON THE ETIOLOGY OF AUTUMN INFANT GASTROENTERITIS

This article deals with further studies on the etiology of a larger series of autumn infant gastroenteritis (AIG) in Beijing in 1979. Fecal samples were collected from 126 and 52 cases in 2 hospitals. The positive rate of rotavirus detected by simple direct electron microscopy (EM) was 38.1To in l hospital and 19.2'70 in the other. Ultrasections were made from fecal sediment of the cases with negative findings by simple direct EM. The positive rate of rotavirus detected by ultrasection EM was 75.670 in 1 hospital and 5270 in the other. The total detec- tion rate obtained by the combined use of these 2 methods reached 84.970 in l hospital. In 1 hospital, adenovirus was found simultaneously with rotavirus in 3 and singly in 2. In the other hospital, adenovirus was found simultaneously with rotavirus in l. Paired sera were obtained from 49 cases, of which 37 ('75.5'/o) showed a 4-fold or higher rise in complement-fixing (CF) antibody to rotavirus, mostly a 32-fold or higher rise. Single convales- cent phase sera were obtained in 6 cases which all gave high rotavirus CF antibody titers. Fecal samples were inoculated into primary thesus monkey kidney monolayer cell cultures for virus isolation. Apart from isolation of a Type 19 ECHO virus strain from l case, addition of trypsin to the fecal samples, maintenance media or both showed no cytopathic changes. A crude extract of a rotavirus particle- positive fecal sample was fed t0 4 newborn pig- lets and one 3-month-old thesus monkey, none of which developed diarrhea. Fecal samples of 133 0f the 178 cases were cultured for bacteria. Only 8 cases (670) showed pathogenic Escherichia coli growth.     

Clinical Study of Poria-Polyporus Anti-Diarrhea Oral Liquor in Treating Infantile Rotavirus Diarrhea

Ｃｌｉｎｉｃａｌ　Ｓｔｕｄｙ　ｏｆ　Ｐｏｒｉａ－Ｐｏｌｙｐｏｒｕｓ　Ａｎｔｉ－Ｄｉａｒｒｈｅａ　Ｏｒａｌ　Ｌｉｑｕｏｒ　ｉｎ　Ｔｒｅａｔｉｎｇ　Ｉｎｆａｎｔｉｌｅ　Ｒｏｔａｖｉｒｕｓ　ＤｉａｒｒｈｅａＷＡＮＧＳｈｕ－ｓｈａｎ（王树山）,ＹＡＮＧＳｏｎ...     

豫北地区婴幼儿腹泻病毒病原学分析

目的了解豫北地区婴幼儿腹泻标本中的轮状病毒、杯状病毒、腺病毒及诺如病毒4种病毒感染的情况。方法采集2009年1月至2009年12月新乡医学院一附院儿科收治的269例小儿腹泻患者粪便标本,采用酶联免疫吸附试验(ELISA)分别检测轮状病毒、杯状病毒、腺病毒、诺如病毒的抗原,并与50例健康儿童的标本相对照。对轮状病毒进行病毒株血清学检测,并采用逆转录聚合酶链式反应(RT-PCR)进行基因分型。结果患儿病毒病原检出率分别为轮状病毒59.2%、杯状病毒7.8%、腺病毒1.1%,未检出诺如病毒;其中混合感染率为2.2%。同时用RT-PCR及ELISA2种方法检查56份粪便标本中轮状病毒,检查一致率达96.43%。56株轮状病毒G血清型分型:血清Ⅰ型3份,Ⅱ型3份,Ⅲ型43份,Ⅸ型4份,未能分型3份。对8株轮状病毒P分型,其中7株为G3P8,1株为G9P8。轮状病毒感染具有明显季节性,以秋、冬季多,杯状病毒以秋季多。结论豫北地区婴幼儿腹泻的病毒病原中以轮状病毒为主,杯状病毒次之,腺病毒检出率低。流行的轮状病毒以G3型为主,G3P8多见。     

婴幼儿肺炎混合感染的病原学研究

婴幼儿肺炎中,肺炎支原体感染及支原体与病毒混合感染也占重要地位。     

Lapine rotavirus: preliminary studies on epizoology and transmission.

Antibody to rotavirus was found in sera of domestic rabbits, wild cottontail rabbits and Snowshoe hares. Eight out of 187 stools from a rabbitry contained rotavirus, though no clinical symptoms were apparent. The virus extracted from one such stool infected a five week old seronegative rabbit and led to distention and congestion of the colon, excretion of virus in the stool and a serological response to virus. Human and Simian rotaviruses did not replicate in the rabbits.