Molecular Epidemiological Analysis of Echovirus 19 Isolated From an Outbreak Associated With Hand, Foot, and Mouth Disease (HFMD) in Shandong Province of China

To elucidate the genetic characterization and molecular epidemiological features of Echovirus 19 （El9） isolates collected from an outbreak associated with hand, foot and mouth disease （HFMD） in Tai＇an city of Shandong Province of China from July to September, 2003. Methods Thirty seven Echovirus 19 isolates were isolated from stool specimens and throat swabs collected during the outbreak, then major capsid （VP1） genomic sequence was determined, and phylogenetic tree was done based on the VP1 sequences among these 37 and other El9 viruses deposited in the Genbank. Also a representative strain named CHN-SD03-TN12 was selected for sequencing of 5′-untranslated regions （5′-UTR）. Results The identity rate was about 98.9%-100% among all these 37 El9 viruses. The genetic relationships between these 37 El9 isolates and other strains reported were also depicted. The identity rate was about 78.4%-78.9% compared with El9 reference strain Burke. The substitutions in the sequence of 5′-UTR resulted in changes in the conjectural properties of 5′-UTR of El9 viruses. Condusion The genetic features of El9 viruses isolated during the outbreak in Shandong Province in 2003 may be associated with a genetic and antigenic drift that changes the virulence of the Shandong isolates, but the molecular changes in Shandong El9 viruses contributing to their phenotype remain to be further illuminated. However, the sequences described in this paper substantiate the changes taken place in capsid VPI and 5′UTR regions. These substitutions may contribute to their tropism and virulence, and play a significant role in pathogenesis and clinical manifestations of the disease.

金葡菌固相抗体特异浓集水中肠道病毒的研究──浓集水样中f_2噬菌体效果观察

采用多聚酶链反应（ＰＣＲ）技术检测水中肠道病毒前，肠道病毒应先行浓集。本研究成功地用金葡菌固相抗体对水中肠道病毒的指标菌ｆ＿２噬菌体进行浓集。（１）金葡菌固相抗体先行混悬于水样中，与ｆ＿２噬菌体结合，然后在３０００ｒ／ｍｉｎ下离心１０ｍｉｎ。１ｍｌ金葡菌固相抗体约可吸附３０００ＰＦＵ以上的ｆ＿２噬菌体。（２）蒸馏水、ＮａＣｌ溶液、磷酸盐缓冲液（ＰＢＳ）及三倍肉汤对金葡菌固相抗体吸附ｆ＿２的效果均无明显影响，吸附率均在９３．７％以上。（３）用ＰＣＲ技术对金葡菌固相抗体捕获的脊髓灰质炎病毒Ｉ型疫苗株（ＰＶＩ）亦可检测。

宁夏地区牛场E种肠道病毒的分离与鉴定

牛肠道病毒感染是近年来国内新报道的一种传染病,其病原体为微RNA病毒科肠道病毒属中的肠道病毒。本研究应用双抗体夹心ELISA方法对宁夏地区的部分牛群肠道病毒感染进行了调查,其感染率为9.85%、27%、28.2%。应用Vero细胞从采自宁夏牛群的粪便样品中分离出2株病毒,采用病毒学技术、过氧化物酶单层细胞试验和分子生物学技术对分离毒鉴定发现,2株病毒均为肠道病毒。应用PCR技术扩增分离毒的部分核苷酸序列,并对其进行比对分析发现,分离的2株病毒均为E种肠道病毒。该结果将为宁夏地区肠道病毒感染的防控提供依据。

河南省牛肠道病毒的遗传变异分析

为了解河南省不同地区牛肠道病毒(BEV)的遗传变异与病毒株间的分子差异,本研究应用PCR方法从河南省不同地区牛场确定为BEV的阳性样品或病毒分离株中扩增了病毒VP1基因并对其进行了序列测定和分析,结果显示检测出的11株BEV中,8株属于E种肠道病毒(EV-E),3株属于F种肠道病毒(EV-F)。VP1基因核苷酸序列的比对分析结果表明,8株不同EV-E毒株间的VP1同源性为70.4%~98.6%,它们与本实验室分离的国内首株EV-E HY12毒株的同源性为69.3%~80.3%;EV-F HeN-A12和HeN-YR91毒株与国内首株EV-F毒株BHM26的同源性为68.6%和70.8%,而与首株山羊EV-F毒株SD-S67的同源性为88.8%和73.7%。进一步分析显示,8株EV-E毒株可以进一步分类为2个基因型EV-E2和EV-E3;而3株EV-F毒株也可进一步分类为2个基因型EV-F1和EV-F4。结果表明,河南省不同地区牛群存在EV-E和EV-F感染和肠道病毒感染的多样性以及病毒株的分子差异,该研究为该病的防控提供流行病学理论依据。