2017年北京市东城区EV-A71和CV-A16分离株VP1基因序列特征分析

目的 通过分析北京市东城区2017年手足口疑似病例样本中肠道病毒A组71(enterovirus 71,EV-A71)和柯萨奇病毒A组16型(coxsackievrius A 16,CV-A16)毒株VP1基因序列特征,为手足口病的防治提供基础资料.方法 利用RT-PCR法对分离得到的EV-A71和CV-A16 VP1序列进行扩增和测序,利用DNAstar5.0和MEGA6.0软件对EV-A71和CV-A16的VP1序列进行比对分析并构建系统发育树.结果 2017年东城区共检测手足口疑似病例咽拭子95份,肠道病毒阳性46份(占48.42％);其中EV-A71阳性4份(占4.21％),CV-A16阳性4份(占4.21％),非EV-A71非CV-A16的其他肠道病毒阳性38份(占40.00％).4株EV-A71分离株核苷酸和氨基酸序列同源性分别为92.89％～99.15％和97.32％～98.66％;3株CV-A16分离株核苷酸和氨基酸序列同源性分别为93.90％～98.76％和98.32％～100.00％.系统进化树显示4株EV-A71属于C4a基因亚型,3株CV-A16属于Blb基因亚型.结论 北京市东城区2017年引起手足口病的病原主要以非EV-A71非CV-A16的其他肠道病毒为主,但是EV-A71和CV-A16仍占一定比例.EV-A71毒株属于C4a基因亚型;CV-A16毒株属于Blb基因亚型;与近年来中国大部分地区毒株的基因分型一致;没有检测到新亚型.

Objective To analyze the clinical and laboratory features of patients with mild and severe HFMD to identify early predictive or diagnostic markers for severe cases. Methods Samples of feces, nasopharyngeal-swab specimens, peripheral blood, serum and cerebral spinal lfuid were collected. Postmortem pathological examination was conducted on 2 dead patients with complication due to neurogenic pulmonary edema. Reverse transcription-polymerase chain-reaction (RT-PCR), culture and isolation of enterovirus 71 (EV71) were performed to detect EV71 infection. Both univariate and multivariate logistic analysis were used to identify factors associated with severe cases. Results EV71 was mainly responsible for HFMD. In this study, 5 isolated EV71 strains belonged to C4 gene subtype. Compared with mild patients, EV71-RNA detection rate was higher and CoxA16 detection rate was lower among severe patients (P < 0.01). Inlfammatory cell inifltration in the lung, cardiac and liver tissues were mild by postmortem pathological examination. It was found that body temperature, vomitting, limb tremor, neutrophil, blood glucose and EV71 infection were significantly related to the severe cases by univariate logisticanalysis. However, after multivariate logistic regression analysis, only vomiting (OR 16.1, CI 2.3-110.5,P < 0.01) and limb tremor (OR 117.6, CI 13.8-1004.5,P < 0.01) were signiifcantly and independently correlated with the severe cases. Conclusions EV71 was mainly responsible for HFMD, particularly for severe cases. Vomiting and limb tremor were predictive markers for severe cases.

肠道病毒71型感染首发肺水肿与肺出血三例报告

Objective To investigate the clinical features of the enterovirus 71 ( EV71 ) infection complicated with pulmonary edema or pulmonary hemorrhage as a fulminant and often fatal illness.Methods The medical records of three cases with EV71 infection were retrospectively reviewed for clinical manifestations, laboratory data, medications, and outcome.Results All the cases were infants and died of the infection. These infants had no skin or mucosal lesions, however, they had sudden onset of cyanosis and tachypnea 1 to 2 days after the onset of the febrile disease with vomiting. All these 3 cases were misdiagnosed and were treated for shock on admission. Pulmonary hemorrhage was not considered in any of the cases on admission. All the cases received tracheal intubation when foamy secretions were discharged from the mouth and nose of the patients and notable cyanosis occurred. After intubation, pink foamy fluid flew out from the endotracheal tube in all the 3 cases. The patients had hyperglycemia and limb weakness, two had tachycardia, and hypertension was found in one case. Chest X-ray showed bilateral or unilateral widespread air space opacity, but the cardiac size and shape were normal. All the patients had leukocytosis. Enterovirus 71 infection was confirmed by detection of specific nucleic acid sequences of the virus from throat swab and tracheal secretions samples and in one case in cerebrospinal fluid.Conclusions Pulmonary edema or pulmonary hemorrhage occurred in the 3 cases with EV71 infection. The initial presentation was nonspecific with fever and vomiting, and sudden appearance of cyanosis, tachypnea, tachycardia, hypertension or hypotension, limb weakness, which may suggest pulmonary edema or hemorrhage. Excessive fluid resuscitation may deteriorate the illness, on the contrary, fluid restriction and inotropic agents, and early intubation with positive pressure mechanical ventilation may be the proper treatment.

肠道病毒71疫苗候选株H3-TY的遗传稳定性研究

了解肠道病毒71(EV71)疫苗候选株H3-TY的遗传稳定性。方法选取EV71疫苗候选株H3-TY种子库中的6个子代病毒，提取RNA，对其VP1基因进行RT-PCR扩增、克隆、序列测定，然后对所获基因序列进行同源性分析并计算同源率。结果H3-TY的6个子代病毒VP1基因之间核苷酸同源性介于99.6％ ～99.8％，氨基酸同源性为100.0％，主要抗原决定簇的氨基酸序列完全一致。结论H3-TY种子库中6代病毒的VP1基因之间高度同源，主要抗原决定簇稳定遗传。

Association of IFNGR2and IL-12B gene polymorphisms with susceptibility to hand,foot and mouth disease infected by EV71