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2015-2016年贵州省手足口病非EV71、非CVA16肠道病毒型别鉴定和分析
Isolation and identification on non-EV71,non-CV-A16 enterovirus of hand-foot-mouth disease in Guizhou,2015-2016
 [PDF]

李俞亭,杨兴林,王芳胜,迟茜文,郭楚娴,魏洪
- , 2018,
Abstract: 目的 探讨2015-2016年贵州省手足口病非EV71(Enterovirus 71,EV 71)、非CVA16(Coxsackievirus A16,CVA16)肠道病毒株的病原构成及优势型别。方法 共收集2015-2016年贵州省儿童手足口病疑似患者肛拭子350例标本,应用实时荧光定量PCR法筛选出非EV71、非CVA16型肠道病毒,采用 RT-PCR法扩增病毒VP1区序列,PCR阳性扩增产物进行测序,对测得序列进行型别鉴定并与国内外各型代表株进行核苷酸同源性分析,构建系统进化树。结果 从收集到的手足口病疑似患者肛拭子标本中,共分离到21株其他肠道病毒,分别为:CVA5、CVA7、CVA9、CVB3(Coxsackievirus B3,CVB3)、 CVB4 、E11(Echovirus11,ECHO11)、 E17各1株,CVA10 、CVB1各2株,CVB5、E7各5株。通过系统发育树揭示,分离到的同型别毒株之间具有同源性,而非同型别的毒株与国内外各代表株之间具有同源性。结论 贵州省引起手足口病的非EV71、非CVA16型肠道病毒型别分布较广,包括CVB组,CVA组和ECHO病毒,其中CVB5、E7为非EV71、非CVA16型肠道病毒主要病原体。
Molecular and seroepidemiologic studies of Enterovirus 71 infection in the State of Pará, Brazil
Castro, Ceyla M.O.;Cruz, Ana Cecília R.;Silva, Edson E. da;Gomes, Maria de Lourdes C.;
Revista do Instituto de Medicina Tropical de S?o Paulo , 2005, DOI: 10.1590/S0036-46652005000200002
Abstract: in many countries, the enterovirus 71 (ev-71) picornaviridae family is associated to hand, foot and mouth disease in addition to acute neurological diseases while in brazil these viruses are more closely associated to the latter group. the aim of this research was to use the first ev-71 isolate of the northern region of brazil in molecular and seroepidemiologic studies. two (2.2%) out of 88 stool samples (44 cases of afp), collected from january 1998 to december 2000 were positive for ev-71 isolation (73442/pa/99). nucleotide sequence of the gen that codifies the vp1 protein showed that isolate 73442/pa/99 was similar to the ev-71 strains belonging to genotype b - more closely identified with ev-71 from north america. neutralization test with 389 sera samples collected from january 1998 to november 2001, from individuals ranging from 0 to 15 years of age living in the city of belém, state of pará showed the following results in relation to isolate 73442/pa/99 and prototype brcr: a total of 207 individuals (53.2%) had neutralization antibodies to both viruses, 167 (42.9%) had no antibodies and 15 showed the presence of neutralizing antibodies to one of the two viruses. only 20.2% of the children aged 0 to 3 had neutralizing antibodies to ev-71, indicating that these children were more susceptible to the infection. both the seroprevalence study and vp1 sequencing were important to demonstrate the spread and the molecular pattern of the ev-71 circulating in the northern region of brazil.
Phenotypic and Genotypic Characteristics of Novel Mouse Cell Line (NIH/3T3)-Adapted Human Enterovirus 71 Strains (EV71:TLLm and EV71:TLLmv)  [PDF]
Carla Bianca Luena Victorio, Yishi Xu, Qimei Ng, Vincent T. K. Chow, Kaw Bing Chua
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0092719
Abstract: Since its identification in 1969, Enterovirus 71 (EV71) has been causing periodic outbreaks of infection in children worldwide and most prominently in the Asia-Pacific Region. Understanding the pathogenesis of Enterovirus 71 (EV71) is hampered by the virus’s inability to infect small animals and replicate in their derived in vitro cultured cells. This manuscript describes the phenotypic and genotypic characteristics of two selected EV71 strains (EV71:TLLm and EV71:TLLmv), which have been adapted to replicate in mouse-derived NIH/3T3 cells, in contrast to the original parental virus which is only able to replicate in primate cell lines. The EV71:TLLm strain exhibited productive infection in all primate and rodent cell lines tested, while EV71:TLLmv exhibited greater preference for mouse cell lines. EV71:TLLmv displayed higher degree of adaptation and temperature adaptability in NIH/3T3 cells than in Vero cells, suggesting much higher fitness in NIH/3T3 cells. In comparison with the parental EV71:BS strain, the adapted strains accumulated multiple adaptive mutations in the genome resulting in amino acid substitutions, most notably in the capsid-encoding region (P1) and viral RNA-dependent RNA polymerase (3D). Two mutations, E167D and L169F, were mapped to the VP1 canyon that binds the SCARB2 receptor on host cells. Another two mutations, S135T and K140I, were located in the VP2 neutralization epitope spanning amino acids 136–150. This is the first report of human EV71 with the ability to productively infect rodent cell lines in vitro.
Enterovirus71 (EV71) Utilise Host microRNAs to Mediate Host Immune System Enhancing Survival during Infection  [PDF]
Yan Long Edmund Lui, Tuan Lin Tan, Wee Hong Woo, Peter Timms, Louise Marie Hafner, Kian Hwa Tan, Eng Lee Tan
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0102997
Abstract: Hand, Foot and Mouth Disease (HFMD) is a self-limiting viral disease that mainly affects infants and children. In contrast with other HFMD causing enteroviruses, Enterovirus71 (EV71) has commonly been associated with severe clinical manifestation leading to death. Currently, due to a lack in understanding of EV71 pathogenesis, there is no antiviral therapeutics for the treatment of HFMD patients. Therefore the need to better understand the mechanism of EV71 pathogenesis is warranted. We have previously reported a human colorectal adenocarcinoma cell line (HT29) based model to study the pathogenesis of EV71. Using this system, we showed that knockdown of DGCR8, an essential cofactor for microRNAs biogenesis resulted in a reduction of EV71 replication. We also demonstrated that there are miRNAs changes during EV71 pathogenesis and EV71 utilise host miRNAs to attenuate antiviral pathways during infection. Together, data from this study provide critical information on the role of miRNAs during EV71 infection.
Developments towards antiviral agents and vaccine against enterovirus 71
EV71感染抗病毒药物及疫苗研究进展

ZHANG Ying-Qiu,YANG Huai-Yi,YANG Zhuo,SHI Ying-Di,ZHAO Chun-Hui,
张迎秋
,杨怀义,杨倬,时迎娣,赵春晖

微生物学通报 , 2012,
Abstract: Hand, foot, and mouth disease (HFMD) outbreaks have been reported in many areas, especially in Aria. Enterovirus 71 (EV71) is the major causative pathogen of HFMD in children and infants, its infection usually accompanied with severe neurological complication which causes high mortality. In recent years, the molecular biology study of EV71 and the progress of antiviral therapies development provide us new strategies for the treatment and prevention of EV71 infection. This paper reviewed the latest achievements in virology and antiviral agents of EV71, including drugs, vaccine, and RNA interference etc, which may be expected to serve as references for related research.
Incidence Rates of Enterovirus 71 Infections in Young Children during a Nationwide Epidemic in Taiwan, 2008–09  [PDF]
Min-Shi Lee ,Pai-Shan Chiang,Shu-Ting Luo,Mei-Liang Huang,Guan-Yuan Liou,Kuo-Chien Tsao,Tzou-Yien Lin
PLOS Neglected Tropical Diseases , 2012, DOI: 10.1371/journal.pntd.0001476
Abstract: Objective Enterovirus 71 (EV71) is causing life-threatening outbreaks in tropical Asia. In Taiwan and other tropical Asian countries, although nationwide EV71 epidemics occur cyclically, age-specific incidence rates of EV71 infections that are critical to estimate disease burden and design vaccine trials are not clear. A nationwide EV71 epidemic occurred in 2008–09 in Taiwan, which provided a unique opportunity to estimate age-specific incidence rates of EV71 infections. Study Design We prospectively recruited 749 healthy neonates and conducted follow-ups from June 2006 to December 2009. Sera were obtained from participants at 0, 6, 12, 24, and 36 months of age for measuring EV71 neutralizing antibody titers. If the participants developed suspected enterovirus illnesses, throat swabs were collected for virus isolation. Results We detected 28 EV71 infections including 20 symptomatic and 8 asymptomatic infections. Age-specific incidence rates of EV71 infection increased from 1.71 per 100 person-years at 0–6 months of age to 4.09, 5.74, and 4.97 per 100 person-years at 7–12, 13–24, and 25–36 months of age, respectively. Cumulative incidence rate was 15.15 per 100 persons by 36 months of age, respectively. Conclusions Risk of EV71 infections in Taiwan increased after 6 months of age during EV71 epidemics. The cumulative incidence rate was 15% by 36 months of age, and 29% of EV71 infections were asymptomatic in young children.
Neutralizing Antibodies to Enterovirus 71 in Belém, Brazil
Gomes, Maria de Lourdes C;Castro, Ceyla Maria O de;Oliveira, Maria José C;Silva, Edson Elias da;
Memórias do Instituto Oswaldo Cruz , 2002, DOI: 10.1590/S0074-02762002000100006
Abstract: non-polio enteroviruses (coxsackievirus a, coxsackievirus b, echovirus and ev 68-72) which belong to the enterovirus (ev) genus, picornaviridae family, may be responsible for acute flaccid paralysis, aseptic meningitis, myocarditis, hepatitis, pleurodinia, neonatal sepsis, hand, foot and mouth disease (hfmd) even though 50-80% of infections are asymptomatic. ev 71 has been responsible for outbreaks and epidemics of hfmd and acute neurologic disease justifying its study in our country. the aim of this study was to detect neutralizing antibodies (ntab) to ev 71 in individuals up to 15 years of age living in belém, state of pará, northern brazil. serum samples from 238 patients attending the virology sector of evandro chagas institute in belém, brazil, were analyzed using microneutralization tests that included rd cells and brcr strain. overall 40.8% (97/238) of tested samples had ntab to ev 71. regarding the distribution per age group, 85.2% (92/108) of patients aged 0-3 years had no ntab to this virus and 69.2% of those 12 to15 years of age were seropositive. these results confirm that ev 71 infection occurs in the city of belém; and that a high rate of individuals in this study were infected aged 3 years and over and, when aged 15 years nearly 70% had ev 71 ntab.
Neutralizing Antibodies to Enterovirus 71 in Belém, Brazil  [cached]
Gomes Maria de Lourdes C,Castro Ceyla Maria O de,Oliveira Maria José C,Silva Edson Elias da
Memórias do Instituto Oswaldo Cruz , 2002,
Abstract: Non-polio enteroviruses (Coxsackievirus A, Coxsackievirus B, Echovirus and EV 68-72) which belong to the enterovirus (EV) genus, Picornaviridae family, may be responsible for acute flaccid paralysis, aseptic meningitis, myocarditis, hepatitis, pleurodinia, neonatal sepsis, hand, foot and mouth disease (HFMD) even though 50-80% of infections are asymptomatic. EV 71 has been responsible for outbreaks and epidemics of HFMD and acute neurologic disease justifying its study in our country. The aim of this study was to detect neutralizing antibodies (NtAb) to EV 71 in individuals up to 15 years of age living in Belém, State of Pará, northern Brazil. Serum samples from 238 patients attending the Virology Sector of Evandro Chagas Institute in Belém, Brazil, were analyzed using microneutralization tests that included RD cells and BrCr strain. Overall 40.8% (97/238) of tested samples had NtAb to EV 71. Regarding the distribution per age group, 85.2% (92/108) of patients aged 0-3 years had no NtAb to this virus and 69.2% of those 12 to15 years of age were seropositive. These results confirm that EV 71 infection occurs in the city of Belém; and that a high rate of individuals in this study were infected aged 3 years and over and, when aged 15 years nearly 70% had EV 71 NtAb.
Inhibition of Enterovirus 71 (EV-71) Infections by a Novel Antiviral Peptide Derived from EV-71 Capsid Protein VP1  [PDF]
Chee Wah Tan, Yoke Fun Chan, Kooi Mow Sim, Eng Lee Tan, Chit Laa Poh
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0034589
Abstract: Enterovirus 71 (EV-71) is the main causative agent of hand, foot and mouth disease (HFMD). In recent years, EV-71 infections were reported to cause high fatalities and severe neurological complications in Asia. Currently, no effective antiviral or vaccine is available to treat or prevent EV-71 infection. In this study, we have discovered a synthetic peptide which could be developed as a potential antiviral for inhibition of EV-71. Ninety five synthetic peptides (15-mers) overlapping the entire EV-71 capsid protein, VP1, were chemically synthesized and tested for antiviral properties against EV-71 in human Rhabdomyosarcoma (RD) cells. One peptide, SP40, was found to significantly reduce cytopathic effects of all representative EV-71 strains from genotypes A, B and C tested, with IC50 values ranging from 6–9.3 μM in RD cells. The in vitro inhibitory effect of SP40 exhibited a dose dependent concentration corresponding to a decrease in infectious viral particles, total viral RNA and the levels of VP1 protein. The antiviral activity of SP40 peptide was not restricted to a specific cell line as inhibition of EV-71 was observed in RD, HeLa, HT-29 and Vero cells. Besides inhibition of EV-71, it also had antiviral activities against CV-A16 and poliovirus type 1 in cell culture. Mechanism of action studies suggested that the SP40 peptide was not virucidal but was able to block viral attachment to the RD cells. Substitutions of arginine and lysine residues with alanine in the SP40 peptide at positions R3A, R4A, K5A and R13A were found to significantly decrease antiviral activities, implying the importance of positively charged amino acids for the antiviral activities. The data demonstrated the potential and feasibility of SP40 as a broad spectrum antiviral agent against EV-71.
Challenges to Licensure of Enterovirus 71 Vaccines  [PDF]
Min-Shi Lee equal contributor,Fan-Chen Tseng equal contributor,Jen-Ren Wang,Chia-Yu Chi,Pele Chong,Ih-Jen Su
PLOS Neglected Tropical Diseases , 2012, DOI: 10.1371/journal.pntd.0001737
Abstract: Human enteroviruses usually cause self-limited infections except polioviruses and enterovirus 71 (EV71), which frequently involve neurological complications. EV71 vaccines are being evaluated in humans. However, several challenges to licensure of EV71 vaccines need to be addressed. Firstly, EV71 and coxsackievirus A (CA) are frequently found to co-circulate and cause hand-foot-mouth disease (HFMD). A polyvalent vaccine that can provide protection against EV71 and prevalent CA are desirable. Secondly, infants are the target population of HFMD vaccines and it would need multi-national efficacy trials to prove clinical protection and speed up the licensure and usage of HFMD vaccines in children. An international network for enterovirus surveillance and clinical trials is urgently needed. Thirdly, EV71 is found to evolve quickly in the past 15 years. Prospective cohort studies are warranted to clarify clinical and epidemiological significances of the antigenic and genetic variations between different EV71 genogroups, which is critical for vaccine design.
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