Identification and molecular typing of Naegleria fowleri from a patient with primary amebic meningoencephalitis in China.

Naegleria fowleri is the only Naegleria spp. known to cause an acute, fulminant, and rapidly fatal central nervous system infection in humans called primary amebic meningoencephalitis (PAM). In 2016, a patient with suspected PAM was found in Zhejiang Province of China. The pathogen was identified by microscopic examination and PCR. The positive PCR products were sequenced and the sequences were aligned using the NCBI BLAST program. The homologous and phylogenetic analysis was conducted using MEGA 6 program. On microscopy of direct smears, motile cells with pseudopodia were observed, and the motion characteristics of the pseudopodia as well as the cell morphology suggested that the pathogens were amoeba trophozoites. Wright-Giemsa-stained smears showed amoeba trophozoites of various shapes, which measured 10-25µm in size; these were characterized by a prominent, centrally placed nucleolus and a vacuolated cytoplasm. PCR was negative for Entamoeba histolytica and Entamoeba dispar, but positive for Naegleria spp. and N. fowleri. The nucleotide sequences acquired in this study have been submitted to GenBank with accession numbers KX909928 and KX909927, respectively. The BLAST analysis revealed that the sequences of KX909928 and KX909927 had 100% similarity with the sequence of the N. fowleri gene (KT375442.1). Sequence alignment and the phylogenetic tree revealed that the N. fowleri collected in this study was classified as genotype 2 and was most closely related to Naegleria lovaniensis. This study confirmed N. fowleri as the agent responsible for the infection in this patient. PAM normally progresses rapidly and is generally universally fatal within a week. Unfortunately this patient died at 2 weeks after the onset of symptoms.

Sero-Molecular Epidemiology of Japanese Encephalitis in Zhejiang, an Eastern Province of China.

BACKGROUND: Sporadic Japanese encephalitis (JE) cases still have been reported in Zhejiang Province in recent years, and concerns about vaccine cross-protection and population-level immunity have been raised off and on within the public health sphere. Genotype I (GI) has replaced GIII as the dominant genotype in Asian countries during the past few decades, which caused considerable concerns about the potential change of epidemiology characteristics and the vaccine effectiveness. The aim of this study was to investigate the prevalence of JE neutralizing antibody and its waning antibody trend after live attenuated JE vaccine immunization. Additionally, this study analyzed the molecular characteristics of the E gene of Zhejiang Japanese encephalitis virus (JEV) strains, and established genetic relationships with other JEV strains. METHODOLOGY/PRINCIPAL FINDINGS: A total of 570 serum specimens were sampled from community population aged from 0 to 92 years old in Xianju county of Zhejiang Province in 2013-2014. Microseroneutralization test results were analyzed to estimate the population immunity and to observe antibody dynamics in vaccinated children. E genes of 28 JEV strains isolated in Zhejiang Province were sequenced for phylogenetic tree construction and molecular characteristics analysis with other selected strains. Positive JE neutralizing antibody rates were higher in residents ≥35 years old (81%~98%) and lower in residents <35 years old (0~57%). 7 or 8 years after the 2nd live attenuated vaccine dose, the antibodies against for 4 different strains with microseroneutralization test were decreased by 55%~73% on seropositive rates and by 25%~38% on GMTs respectively. JEV strains isolated in recent years were all grouped into GI, while those isolated in the 1980s belonged to GIII. On important amino acid sites related to antigenicity, there was no divergence between the Zhejiang JE virus strains and the vaccine strain (SA14-14-2). CONCLUSION/SIGNIFICANCES: JE neutralizing antibody positive rates increase in age ≥10 years old population, likely reflecting natural infection or natural boosting of immunity through exposure to wild virus. JE seropositivity rates were quite low in <35 years old age groups in Zhejiang Province. Waning of neutralizing antibody after live attenuated vaccine immunization was observed, but the clinical significance should be further investigated. Both the peripheral antibody response and genetic characterization indicate that current live attenuated JE vaccine conferred equal neutralizing potency against GI or GIII of wild strains. GI has replaced GIII as the dominant genotype in Zhejiang in the past few decades. Although the chance of exposure to wild JE virus has reduced, the virus still circulates in nature; therefore, it is necessary to implement immunization program for children continually and to conduct surveillance activity periodically.

A hospital-associated measles outbreak among individuals not targeted for vaccination in eastern China, 2014.

BACKGROUND: An outbreak of measles occurred in early 2014 among individuals not targeted for vaccination-adults, and infants too young to vaccinate, in Xiangshan County, Zhejiang Province, in eastern China. OBJECTIVE: We conducted an investigation to identify risk factors responsible for this outbreak and to provide evidence-based recommendations for measles elimination strategies in China. METHODS: Measles was diagnosed using national standard case definitions. In a case-control study, 20 randomly selected measles patients were matched with controls selected from the same village or community as each case in a 1:2 case-to-control ratio. Controls were matched on age, within 5 years, and gender. We compared exposure histories during the 7-21 days before rash onset of the case and the same time period for the matched controls. We also conducted a measles antibody seroprevalence survey of a convenient sample of residual serum obtained from healthy patients during routine care in a hospital. RESULTS: The outbreak consisted of 45 measles cases, with an attack rate of 8.9/100,000 total population. Among cases, 91.1% (41/45) were adults (ranged 23-51 years) who had unknown vaccination histories; the other cases were infants younger than 8 months of age. The case-control study showed major risk factors to be a visit to Hospital X (OR(MH)=7.3, 95% CI: 1.8-30.7) and treatment in an IV room in Hospital X (OR(MH)=11.0, 95% CI: 1.3-96.1). The seroprevalence survey showed that 88.8% of adults had measles IgG antibodies, and that 100% of children 2-19 years of age were seropositive. CONCLUSIONS: The outbreak was primarily among age groups not targeted for vaccination-primarily adults, but with some children too young to vaccinate. Visiting a hospital was the major risk factor for measles transmission. We conclude that in addition to maintaining high 2-dose coverage with measles vaccine, working with hospital infection control programs to implement evidence-based strategies to prevent or limit hospital transmission is an important action for eliminating measles in eastern China.

[Self-controlled case-series (SCCS) method as a tool for the evaluation on the safety of vaccine].

INTRODUCTION: Safety on vaccine issues has been under greater concern. Epidemiologically, classical study designs on investigating the association between a rare adverse event and vaccine inoculation usually confronted with lower feasibility in practice. Self-controlled case series (SCCS) method was derived from cohort studies. The key advantage of this method is that it only uses data related to cases and trying to find relative incidence of events in the 'at risk' periods relative to the 'controlled' periods. A further benefit of this method is that all the fixed confounders are controlled implicitly, by self-control, thus provides high statistic powers. With these advantages, SCCS is suitable for the causality assessment on rare but severe adverse events caused by immunization, which has been widely used,abroad. However,the methodology of SCCS is still being developed,and the areas in use have been expanded to the studies related to the safety and effectiveness of drugs, efficacy of vaccines as well as risk factors of disease.

[Evaluation of the effect of varicella outbreak control measures through a discrete time delay SEIR model].

OBJECTIVE: forecast the epidemic trend and to evaluate the effect of outbreak control measures by investigation of a varicella outbreak event with a discrete time delay SEIR model. METHODS: A discrete time delay model was formulated by discretization method based on a continuous SEIR model with the consideration of the time delay effect on latent period and communicable period. The epidemic trend forecast was carried out based on the number of expected cases. The theoretical effect evaluation was assessed by comparing the results from different emergency control measures. RESULTS: Without any control measures, the theoretical attack rate was 30.16% (504/1671). The course of the epidemic lasted for 4 months and the peak epidemic time was 78 days after the onset of the first case. 'Generation' phenomenon had been observed in the course of the epidemic with the interval of two weeks. The actual number of cases was decreased by 89.48% (451/504) compared with the number of expected cases under no control measure scenario. With the rigorous quarantine measure for all cases on their onset day, when the measure was conducted on 0, 14, 28, 42 days after the onset of the first case, the total numbers of expected cases were 22, 59, 127 and 220 respectively. With the quarantine measure conducted on 14 days after the onset of the first case, when the proportion of quarantine was 30%, 50%, 70%, 90%, the total number of expected cases were 485, 457, 386 and 169, respectively. With the emergent vaccination for all persons, when the measure was conducted on 0, 14, 28, 42 days after the onset of the first case, the total numbers of expected cases were 7, 26, 81 and 202 respectively. With the emergent vaccination conducted on 14 days after the onset of the first case, when the immunization coverage rate was 30%, 50%, 70%, 90%, the total numbers of expected cases were 354, 246, 127 and 40, respectively. CONCLUSION: The number of expected cases can be regarded as the reference to evaluate the effect of control measures. The simulation results suggest that it will get more benefits to conduct control measures earlier in varicella outbreak events, and the effect of emergent vaccination is better than that of quarantine measure under the same conditions.

[Design of field studies on vaccines which involves capturing indirect effect].

For most variety of vaccines, two types of effects-direct and indirect, can result from immunization programs. Unimmunized individuals in the population that receive immunization program can benefit from the vaccines in addition to the protection from immunization. The classical vaccine trails allocate individuals into study and control arms with individual randomization, so the programs' cost-benefit is underestimated due to the impossible measurement on indirect effect. The objectives of the present work are to introduce the conceptual framework, developed by Halloran et al, for four types of study designs that differentiate and account for direct, indirect, total and overall effects of intervention programs, and to explain the relationships of these effects as well. With the examples of field trails of oral cholera vaccines in Bangladesh and Vi typhoid vaccine in India, further issues refer to the identification of indirect effect. The application in practice is also discussed.

[Bootstrap method-based estimation on the confidence interval for additive interaction in cohort studies].

Interaction assessment is an important step in epidemiological analysis. When etiological study is carried out, the logarithmic models such as logistic model or Cox proportional hazard model are commonly used to estimate the independent effects of the risk factors. However, estimating interaction between risk factors by the regression coefficient of the product term is on multiplicative scale, and for public-health purposes, it is supposed to be on additive scale or departure from additivity. This paper illustrates with a example of cohort study by fitting Cox proportional hazard model to estimate three measures for additive interaction which presented by Rothman. Adopting the S-Plus application with a built-in Bootstrap function, it is convenient to estimate the confidence interval for additive interaction. Furthermore, this method can avoid the exaggerated estimation by using ORs in a cohort study to gain better precision. When using the complex combination models between additive interaction and multiplicative interaction, it is reasonable to choose the former one when the result is inconsistent.