Safety and immunogenicity of a broad-spectrum mosaic vaccine as a booster dose against SARS-CoV-2 Omicron and other circulating variants

BACKGROUNDThe rising breakthrough infections caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, especially Omicron and its sub-lineages, have raised an urgent need to develop broad-spectrum vaccines against coronavirus disease 2019 (COVID-19). We have developed a mosaic-type recombinant vaccine candidate, named NVSI-06-09, having immune potentials against a broad range of SARS-CoV-2 variants. METHODSAn ongoing randomized, double-blind, controlled phase 2 trial was conducted to evaluate the safety and immunogenicity of NVSI-06-09 as a booster dose in subjects aged 18 years and older from the United Arab Emirates (UAE), who had completed two or three doses of BBIBP-CorV vaccinations at least 6 months prior to the enrollment. The participants were randomly assigned with 1:1 to receive a booster dose of NVSI-06-09 or BBIBP-CorV. The primary outcomes were immunogenicity and safety against SARS-CoV-2 Omicron variant, and the exploratory outcome was cross-immunogenicity against other circulating strains. RESULTSA total of 516 participants received booster vaccination. Interim results showed a similar safety profile between NVSI-06-09 and BBIBP-CorV booster groups, with low incidence of adverse reactions of grade 1 or 2. For immunogenicity, by day 14 after the booster vaccination, the fold rises in neutralizing antibody geometric mean titers (GMTs) from baseline level elicited by NVSI-06-09 were remarkably higher than those by BBIBP-CorV against the prototype strain (19.67 vs 4.47-fold), Omicron BA.1.1 (42.35 vs 3.78-fold), BA.2 (25.09 vs 2.91-fold), BA.4 (22.42 vs 2.69-fold), and BA.5 variants (27.06 vs 4.73-fold). Similarly, the neutralizing GMTs boosted by NVSI-06-09 against Beta and Delta variants were also 6.60-fold and 7.17-fold higher than those boosted by BBIBP-CorV. CONCLUSIONSA booster dose of NVSI-06-09 was well-tolerated and elicited broad-spectrum neutralizing responses against SARS-CoV-2 prototype strain and immune-evasive variants, including Omicron and its sub-lineages. The immunogenicity of NVSI-06-09 as a booster vaccine was superior to that of BBIBP-CorV. (Funded by LIBP and BIBP of Sinopharm; ClinicalTrials.gov number, NCT05293548).

Safety and immunogenicity of a hybrid-type vaccine booster in BBIBP-CorV recipients: a randomized controlled phase 2 trial

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with immune escape ability raises the urgent need for developing cross-neutralizing vaccines against the virus. NVSI-06-08 is a potential broad-spectrum recombinant COVID-19 vaccine that integrates the antigens from multiple SARS-CoV-2 strains into a single immunogen. Here, we evaluated the safety and immunogenicity of NVSI-06-08 as a heterologous booster dose in adults previously vaccinated with the inactivated vaccine BBIBP-CorV in a randomized, double-blind, controlled, phase 2 trial conducted in the United Arab Emirates (NCT05069129). Three groups of healthy adults over 18 years of age (600 participants per group) who had administered two doses of BBIBP-CorV 4-6-month, 7-9-month and >9-month earlier, respectively, were vaccinated with either a homologous booster of BBIBP-CorV or a heterologous booster of NVSI-06-08. The primary outcome was immunogenicity and safety of booster vaccinations. The exploratory outcome was cross-reactive immunogenicity against multiple SARS-CoV-2 variants of concerns (VOCs). The incidence of adverse reactions was low in both booster vaccinations, and the overall safety profile of heterologous boost was quite similar to that of homologous boost. Heterologous NVSI-06-08 booster was immunogenically superior to homologous booster of BBIBP-CorV. Both Neutralizing and IgG antibodies elicited by NVSI-06-08 booster were significantly higher than by the booster of BBIBP-CorV against not only SARS-CoV-2 prototype strain but also multiple VOCs. Especially, the neutralizing activity induced by NVSI-06-08 booster against the immune-evasive Beta variant was no less than that against the prototype strain, and a considerable level of neutralizing antibodies against Omicron (GMT: 367.67; 95%CI, 295.50-457.47) was induced by heterologous booster, which was substantially higher than that boosted by BBIBP-CorV (GMT: 45.03; 95%CI, 36.37-55.74). Our findings showed that NVSI-06-08 was safe and immunogenic as a booster dose following two doses of BBIBP-CorV, which was immunogenically superior to homologous boost with another dose of BBIBP-CorV. Our study also indicated that the design of hybrid antigen may provide an effective strategy for broad-spectrum vaccine developments.

Safety and immunogenicity of a heterologous boost with a recombinant vaccine, NVSI-06-07, in the inactivated vaccine recipients from UAE: a phase 2 randomised, double-blinded, controlled clinical trial

BackgroundThe increased coronavirus disease 2019 (COVID-19) breakthrough cases pose the need of booster vaccinations. In this study, we reported the safety and immunogenicity of a heterologous boost with a recombinant COVID-19 vaccine (CHO cells), named NVSI-06-07, as a third dose in participants who have previously received two doses of the inactivated vaccine (BBIBP-CorV) at pre-specified time intervals. Using homologous boost with BBIBP-CorV as control, the safety and immunogenicity of the heterologous boost with NVSI-06-07 against various SARS-CoV-2 strains, including Omicron, were characterized. MethodsThis study is a single-center, randomised, double-blinded, controlled phase 2 trial for heterologous boost of NVSI-06-07 in BBIBP-CorV recipients from the United Arab Emirates (UAE). Healthy adults (aged [≥]18 years) were enrolled and grouped by the specified prior vaccination interval of BBIBP-CorV, i.e., 1-3 months, 4-6 months or [≥]6 months, respectively, with 600 individuals per group. For each group, participants were randomly assigned at 1:1 ratio to receive either a heterologous boost of NVSI-06-07 or a homologous booster dose of BBIBP-CorV. The primary outcome was to comparatively assess the immunogenicity between heterologous and homologous boosts at 14 and 28 days post-boosting immunization, by evaluation of the geometric mean titers (GMTs) of IgG and neutralizing antibodies as well as the corresponding seroconversion rate ([≥]4-fold rise in antibody titers). The secondary outcomes were the safety profile of the boosting strategies within 30 days post vaccination. The exploratory outcome was the immune efficacy against Omicron and other variants of concern (VOCs) of SARS-CoV-2. This trial is registered with ClinicalTrials.gov, NCT05033847. FindingsA total of 1800 individuals who have received two doses of BBIBP-CorV were enrolled, of which 899 participants received a heterologous boost of NVSI-06-07 and 901 received a homologous boost for comparison. No vaccine-related serious adverse event (SAE) and no adverse events of special interest (AESI) were reported. 184 (20{middle dot}47%) participants in the heterologous boost groups and 177 (19{middle dot}64%) in the homologous boost groups reported at least one adverse reaction within 30 days. Most of the local and systemic adverse reactions reported were grades 1 (mild) or 2 (moderate), and there was no significant difference in the overall safety between heterologous and homologous boosts. Immunogenicity assays showed that the seroconversion rates in neutralizing antibodies against prototype SARS-CoV-2 elicited by heterologous boost were 89{middle dot}96% - 97{middle dot}52% on day 28 post-boosting vaccination, which was much higher than what was induced by homologous boost (36{middle dot}80% - 81{middle dot}75%). Similarly, in heterologous NVSI-06-07 booster groups, the neutralizing geometric mean titers (GMTs) against the prototype strain increased by 21{middle dot}01 - 63{middle dot}85 folds from baseline to 28 days post-boosting vaccination, whereas only 4{middle dot}20 - 16{middle dot}78 folds of increases were observed in homologous BBIBP-CorV booster group. For Omicron variant, the neutralizing antibody GMT elicited by the homologous boost of BBIBP-CorV was 37{middle dot}91 (95%CI, 30{middle dot}35-47{middle dot}35), however, a significantly higher level of neutralizing antibodies with GMT 292{middle dot}53 (95%CI, 222{middle dot}81-384{middle dot}07) was induced by the heterologous boost of NVSI-06-07, suggesting that it may serve as an effective boosting strategy combating the pandemic of Omicron. The similar results were obtained for other VOCs, including Alpha, Beta and Delta, in which the neutralizing response elicited by the heterologous boost was also significantly greater than that of the homologous boost. In the participants primed with BBIBP-CorV over 6 months, the largest increase in the neutralizing GMTs was obtained both in the heterologous and homologous boost groups, and thus the booster vaccination with over 6 months intervals was optimal. InterpretationOur findings indicated that the heterologous boost with NVSI-06-07 was safe, well-tolerated and immunogenic in adults primed with a full regimen of BBIBP-CorV. Compared to homologous boost with a third dose of BBIBP-CorV, incremental increases in immune responses were achieved by the heterologous boost with NVSI-06-07 against SARS-CoV-2 prototype strain, Omicron variant, and other VOCs. The heterologous BBIBP-CorV/NVSI-06-07 prime-boosting vaccination may be valuable in preventing the pandemic of Omicron. The optimal booster strategy was the heterologous boost with NVSI-06-07 over 6 months after a priming with two doses of BBIBP-CorV. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSWe searched PubMed for clinical trials or prospective/cohort studies involving heterologous booster vaccination in non-immunocompromised population published up to Dec 25, 2021, using the term "(COVID) AND (vaccin*) AND (clinical trial OR cohort OR prospective) AND (heterologous) AND (booster OR prime-boost OR third dose)" with no language restrictions. Nine studies of heterologous prime-boost vaccinations with adenovirus-vector vaccines (ChAdOx1 nCov-19, Oxford-AstraZeneca, Ad26.COV2.S, Janssen) and mRNA vaccines (BNT162b2, Pfizer-BioNtech; mRNA1273, Moderna) were identified. The adenovirus-vector and mRNA heterologous prime-boost vaccination was found to be well tolerated and immunogenic. In individuals primed with adenovirus-vector vaccine, mRNA booster vaccination led to greater immune response than homologous boost. However, varied results were obtained on whether heterologous boost was immunogenically superior to the homologous mRNA prime-boost vaccination. Besides that, A preprint trial in population previously immunized with inactivated vaccines (CoronaVac, Sinovac Biotech) showed that the heterologous boost with adenovirus-vector vaccine (Convidecia, CanSino Biologicals) was safe and induced higher level of live-virus neutralizing antibodies than by the homogeneous boost. A pilot study reported that boosting with BNT162b2 in individuals primed with two doses of inactivated vaccines (BBIBP-CorV) was significantly more immunogenic than homologous vaccination with two-dose of BNT162b2. In addition, a preprint paper demonstrated that heterologous boost of ZF2001, a recombinant protein subunit vaccine, after CoronaVac or BBIBP-CorV vaccination potently improved the immunogenicity. But only a small size of samples was tested in this study and the live-virus neutralization was not detected. Till now, it is still lacking a formal clinical trial to evaluate the immunogenicity and safety of the heterologous prime-boost vaccination with an inactivated vaccine followed by a recombinant protein subunit-based vaccine. Added value of this studyTo our knowledge, this is the first reported result of a large-scale randomised, controlled clinical trial of heterologous prime-boost vaccination with an inactivated vaccine followed by a recombinant protein subunit vaccine. This trial demonstrated that the heterologous prime-booster vaccination with BBIBP-CorV/NVSI-06-07 is safe and immunogenic. Its immunoreactivity is similar to that of homologous vaccination with BBIBP-CorV. Compared to homologous boost, heterologous boost with NVSI-06-07 in BBIBP-CorV recipients elicited significantly higher immunogenicity not only against the SARS-CoV-2 prototype strain but also against Omicron and other variants of concern (VOCs). Implications of all the available evidenceBooster vaccination is considered an effective strategy to improve the protection efficacy of COVID-19 vaccines and control the epidemic waves of SARS-CoV-2. Data from our trial suggested that the booster vaccination of NVSI-06-07 in BBIBP-CorV recipients significantly improved the immune responses against various SARS-CoV-2 strains, including Omicron. Due to no Omicron-specific vaccine available currently, the BBIBP-CorV/NVSI-06-07 heterologous prime-boost might serve as an effective strategy combating Omicron variant. Besides that, BBIBP-CorV has been widely inoculated in population, and thus further boosting vaccination with NVSI-06-07 is valuable in preventing the COVID-19 pandemic. But further studies are needed to assess the long-term protection of BBIBP-CorV/NVSI-06-07 prime-booster vaccination.

CD147 antibody specifically and effectively inhibits infection and cytokine storm of SARS-CoV-2 variants

SARS-CoV-2 and its variants are raging worldwide. Unfortunately, the global vaccination is not efficient enough to attain a vaccine-based herd-immunity and yet no special and effective drug is developed to contain the spread of the disease. Previously we have identified CD147 as a novel receptor for SARS-CoV-2 infection. Here, we demonstrated that CD147 antibody effectively inhibits infection and cytokine storm caused by SARS-CoV-2 variants. In CD147KO VeroE6 cells, infections of SARS-CoV-2, its variants (B.1.1.7, B.1.351) and pseudovirus mutants (B.1.1.7, B.1.351, B.1.525, B.1.526 (S477N), B.1.526 (E484K), P.1, P.2, B.1.617.1, B.1.617.2) were decreased. Meanwhile, CD147 antibody effectively blocked the entry of variants and pseudomutants in VeroE6 cells, and inhibited the expression of cytokines. A model of SARS-CoV-2-infected hCD147 transgenic mice was constructed, which recapitulated the features of exudative diffuse alveolar damage and dynamic immune responses of COVID-19. CD147 antibody could effectively clear the virus and alveolar exudation, resolving the pneumonia. We found the elevated level of cyclophilin A (CyPA) in plasma of severe/critical cases, and identified CyPA as the most important proinflammatory intermediate causing cytokine storm. Mechanistically, spike protein of SARS-CoV-2 bound to CD147 and initiated the JAK-STAT pathway, which induced expression of CyPA. CyPA reciprocally bound to CD147, triggered MAPK pathway and consequently mediated the expression of cytokine and chemokine. In conclusion, CD147 is a critical target for SARS-CoV-2 variants and CD147 antibody is a promising drug to control the new wave of COVID-19 epidemic.

Landscapes and dynamic diversifications of B-cell receptor repertoires in COVID-19 patients

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the pandemic of coronavirus disease 2019 (COVID-19). Great international efforts have been put into the development of prophylactic vaccines and neutralizing antibodies. However, the knowledge about the B cell immune response induced by the SARS-CoV-2 virus is still limited. Here, we report a comprehensive characterization of the dynamics of immunoglobin heavy chain (IGH) repertoire in COVID-19 patients. By using next-generation sequencing technology, we examined the temporal changes in the landscape of the patients immunological status, and found dramatic changes in the IGH within the patients immune system after the onset of COVID-19 symptoms. Although different patients have distinct immune responses to SARS-CoV-2 infection, by employing clonotype overlap, lineage expansion and clonotype network analyses, we observed a higher clonotype overlap and substantial lineage expansion of B cell clones during 2-3 weeks of illness, which is of great importance to B-cell immune responses. Meanwhile, for preferences of V gene usage during SARS-CoV-2 infection, IGHV3-74 and IGHV4-34 and IGHV4-39 in COVID-19 patients were more abundant than that of healthy controls. Overall, we present an immunological resource for SARS-CoV-2 that could promote both therapeutic development as well as mechanistic research.

Recombinant SARS-CoV-2 RBD molecule with a T helper epitope as a built in adjuvant induces strong neutralization antibody response

Without approved vaccines and specific treatment, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is spreading around the world with above 20 million COVID-19 cases and approximately 700 thousand deaths until now. An efficacious and affordable vaccine is urgently needed. The Val308 - Gly548 of Spike protein of SARS-CoV-2 linked with Gln830 - Glu843 of Tetanus toxoid (TT peptide) (designated as S1-4) and without TT peptide (designated as S1-5), and prokaryotic expression, chromatography purification and the rational renaturation of the protein were performed. The antigenicity and immunogenicity of S1-4 protein was evaluated by Western Blotting (WB) in vitro and immune responses in mice, respectively. The protective efficiency of it was measured by virus neutralization test in Vero E6 cells with SARS-CoV-2. S1-4 protein was prepared to high homogeneity and purity by prokaryotic expression and chromatography purification. Adjuvanted with Alum, S1-4 protein stimulated a strong antibody response in immunized mice and caused a major Th2-type cellular immunity compared with S1-5 protein. Furthermore, the immunized sera could protect the Vero E6 cells from SARS-CoV-2 infection with neutralization antibody GMT 256. The candidate subunit vaccine molecule could stimulate strong humoral and Th1 and Th2-type cellular immune response in mice, giving us solid evidence that S1-4 protein could be a promising subunit vaccine candidate.

A non-competing pair of human neutralizing antibodies block COVID-19 virus binding to its receptor ACE2

Neutralizing antibodies could be antivirals against COVID-19 pandemics. Here, we report the isolation of four human-origin monoclonal antibodies from a convalescent patient in China. All of these isolated antibodies display neutralization abilities in vitro. Two of them (B38 and H4) block the binding between RBD and vial cellular receptor ACE2. Further competition assay indicates that B38 and H4 recognize different epitopes on the RBD, which is ideal for a virus-targeting mAb-pair to avoid immune escape in the future clinical applications. Moreover, therapeutic study on the mouse model validated that these two antibodies can reduce virus titers in the infected mouse lungs. Structure of RBD-B38 complex revealed that most residues on the epitope are overlapped with the RBD-ACE2 binding interface, which explained the blocking efficacy and neutralizing capacity. Our results highlight the promise of antibody-based therapeutics and provide the structural basis of rational vaccine design. One Sentence SummaryA pair of human neutralizing monoclonal antibodies against COVID-19 compete cellular receptor binding but with different epitopes, and with post-exposure viral load reduction activity.

Mutations, Recombination and Insertion in the Evolution of 2019-nCoV

BackgroundThe 2019 novel coronavirus (2019-nCoV or SARS-CoV-2) has spread more rapidly than any other betacoronavirus including SARS-CoV and MERS-CoV. However, the mechanisms responsible for infection and molecular evolution of this virus remained unclear. MethodsWe collected and analyzed 120 genomic sequences of 2019-nCoV including 11 novel genomes from patients in China. Through comprehensive analysis of the available genome sequences of 2019-nCoV strains, we have tracked multiple inheritable SNPs and determined the evolution of 2019-nCoV relative to other coronaviruses. ResultsSystematic analysis of 120 genomic sequences of 2019-nCoV revealed co-circulation of two genetic subgroups with distinct SNPs markers, which can be used to trace the 2019-nCoV spreading pathways to different regions and countries. Although 2019-nCoV, human and bat SARS-CoV share high homologous in overall genome structures, they evolved into two distinct groups with different receptor entry specificities through potential recombination in the receptor binding regions. In addition, 2019-nCoV has a unique four amino acid insertion between S1 and S2 domains of the spike protein, which created a potential furin or TMPRSS2 cleavage site. ConclusionsOur studies provided comprehensive insights into the evolution and spread of the 2019-nCoV. Our results provided evidence suggesting that 2019-nCoV may increase its infectivity through the receptor binding domain recombination and a cleavage site insertion. One Sentence SummaryNovel 2019-nCoV sequences revealed the evolution and specificity of betacoronavirus with possible mechanisms of enhanced infectivity.

The Pathogenicity of 2019 Novel Coronavirus in hACE2 Transgenic Mice

Severe acute respiratory syndrome CoV-2 (SARS-CoV-2) caused the Corona Virus Disease 2019 (COVID-19) cases in China has become a public health emergency of international concern (PHEIC). Based on angiotensin converting enzyme 2 (ACE2) as cell entry receptor of SARS-CoV, we used the hACE2 transgenic mice infected with SARS-CoV-2 to study the pathogenicity of the virus. Weight loss and virus replication in lung were observed in hACE2 mice infected with SARS-CoV-2. The typical histopathology was interstitial pneumonia with infiltration of significant lymphocytes and monocytes in alveolar interstitium, and accumulation of macrophages in alveolar cavities. Viral antigens were observed in the bronchial epithelial cells, alveolar macrophages and alveolar epithelia. The phenomenon was not found in wild type mice with SARS-CoV-2 infection. The pathogenicity of SARS-CoV-2 in hACE2 mice was clarified and the Kochs postulates were fulfilled as well, and the mouse model may facilitate the development of therapeutics and vaccines against SARS-CoV-2.

Comparison of technical specifications between domestic and foreign high level isolated units / 中华实验和临床病毒学杂志

Objective@#By analysis the differences of technical specifications among National Health Commission of the People’s Republic of China, Centers for Disease Control and Prevention (CDC) and European Network of Infectious Disease (EUNID)domestic and foreign, to provide a reference for the construction of high-level isolation units (HLIUs) in China.

Impact of sealing method and layer number of autoclave bag on autoclave sterilization effect in the BSL-3 laboratory / 中华实验和临床病毒学杂志

Objective@#To evaluate, the method of different packing layer and sealing mode for the packaging of the infectious wastes in order to achieve better autoclave sterilization effect.@*Methods@#In the autoclave cycles P4 and P9, different packing layer (single, double and triple) and sealing mode (slightly folded, autoclave tape, nylon tape closure) were applied to the internal infectious waste sterilization, biological indicators were used to detect the effect of autoclave sterilization of the upper, middle and lower layers, respectively.@*Results@#1) In the autoclave cycle P4, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively in the upper layer 100%, 0%, 0%, in the middle layer 67%, 20%, 0%, and in the lower layer was 0%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, the middle layer and the lower layer were respectively the in the upper layer 100%, 85%, 0%, the middle layer 45%, 0%, 0%, and the lower layer 10%, 0%, 0%. the single autoclave bag packaging sealing method wa based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer 100%, 100%, 0%, the middle layer 70%, 45%, 0%, and the lower layer 12.5%, 0%, 0%. (2) In the autoclave cycle P9, the triple autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper layer, middle and lower layers were respectively the upper layer all 100%, the middle layer 100%, 100%, 70%, and the lower layer 100%, 100%, 30%. The double autoclave bags packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were respectively the upper layer all 100%, the middle layer all 100% and the lower layer is 100%, 100%, and 60%. The single autoclave bag packaging sealing method was based on the slight folding, autoclave tape, and nylon taple, the biological indicator negative rate of the internal infectious waste in the upper, middle and lower layers were all 100%.@*Conclusions@#When autoclave is recommended for sterilization of infectious waste, the best effect was achieved with a single unsealed autoclave bag. If it is necessary to seal, the autoclave bag should not be completely sealed, avoiding incomplete penetration of steam into the interior space, otherwise the infectious waste can not be completely sterilized.

Pathogenic composition of hand, foot and mouth disease and enterovirus-A71 and coxsackievirus-A16 genetic characteristics analysis in Ningxia, 2016-2017 / 中华实验和临床病毒学杂志

Objective@#To understand the pathogenic composition of hand, foot and mouth disease (HFMD) in Ningxia Hui Autonomous Region (Ningxia) from 2016 to 2017, and analyze the genetic characteristics of the main pathogens enterovirus (EV)-A71 and coxsackievirus (CV)-A16.@*Methods@#Analysis of the result of nucleic acid testing of HFMD in Ningxia from 2016 to 2017 to determine the pathogenic composition of HFMD. The complete VP1 coding region was amplified by RT-PCR and the gene sequence was determined for the enterovirus strains sent to the National HFMD Network Monitoring Laboratory in Ningxia from 2016 to 2017. BLAST analysis confirmed the serotype of the strain, and the phylogenetic tree was constructed respectively by selecting EV-A71 and CV-A16 isolates.@*Results@#The leading pathogens of HFMD in Ningxia of 2016 and 2017 were other EV (397, 43.72%) and EV-A71 (918, 56.18%) respectively, and the dominant pathogens in different months may differ. The pathogenic composition causing HFMD in the past two years has changed from CV-A16 and other EV to EV-A71 and other EV. The isolated EV-A71 strains were C4a evolutionary branch and the isolated CV-A16 strains were B1b evolutionary branch.@*Conclusions@#Compared to 2016, in 2017 EV-A71, CV-A16 and other EV changed dynamically. Dynamic monitoring of EV-A71 in Ningxia is of great significance to guide the strategy of using EV-A71 vaccine, concentrating medical resources to strengthen the treatment and reduce the mortality rate of severe HFMD cases.

Analysis on global Rhinovirus research advances / 中华实验和临床病毒学杂志

Objective@#To explore the research status of rhinovirus (RV) through analysis of rhinovirus literature using GoPubMed.@*Methods@#"Rhinovirus" was used as the major subject word and the rhinovirus literature was collected at PubMed database (from Jan 1, 1970 to April 16, 2018). The high frequency subject words of rhinovirus related literature and the distribution of countries, cities, and journals were analyzed through a bibliometrical analysis method .@*Results@#A total of 5 367 reports were retrieved from PubMed. The quantity of rhinovirus papers increased overall year by year. The highest number of papers were mainly published in developed countries. The highest number of papers on RV were mainly published in J Virol among all journals related with rhinovirus and Tyrrell D published the highest number of papers in all authors contributed to articles on rhinovirus. The rhinovirus, human, virus, respiratory tract infection were the high frequency subject words in the rhinovirus research.@*Conclusions@#Rhinovirus research is becoming one of research hotspots according to the statistical analysis of the research literature on rhinoviruses by GoPubMed.

A study of establishment of non-standard methods in qualification accreditation of virological laboratories / 中华实验和临床病毒学杂志

The laboratories of National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, have developed a series of experimental method . These method have unique advantages over the national standard method and industry standard method . How to make these method become public products through legal procedures to serve disease prevention and control more extensively is an obligatory task for national virological laboratories. This article explores the establishment, verification, validation, and examination of virological non-standard method under laboratory certification and accreditation conditions through empirical research on the " RT-RAA method for detecting EV71/CA16 and other viruses" .

Consideration on microbiological laboratory biosafety of China in the new era / 中华实验和临床病毒学杂志

As one part of Overall National Security, Biosafety has became a basic strategy for adhering to and developing the socialism with Chinese characteristics in the new era, which also meets the requirements of national strategies. Since 2004, microbiological laboratory biosafety of China has made significant progress in China, and gave a positive contribution to Ebola fight in Serra Leone in 2014. In the new era, microbiological laboratory biosafety of China should insist on the core problem, cover the total issues and make sure enough measures be in place. Under the principal of national strategy of Implementing Healthy China, the Belt and the Road Initiative, building a community of human destiny, microbiological laboratory biosafety will centre on "3B" problems related to biosafety, biosecurity and biotechnology, and provide necessary guarantee to re-emerging infectious disease control and public health security.

Establishment of a real-time PCR method to identify Ekpoma virus gene in blood sample of a returnee from Angola / 中华实验和临床病毒学杂志

Objective@#To establish quantitative real-time PCR (qPCR) method based on Taqman probe for detecting Ekpoma virus (EKV).@*Methods@#According to the conserved region of gene in EKV genome from GenBank, primers and probe for qPCR were designed. Validity and sensitivity were evaluated in this study. Both whole blood and serum of a returnee from Angola were tested by the established EKV-1 and EKV-2 qPCR method .@*Results@#Sensitivity of EKV-1 and EKV-2 qPCR method was respectively 41 copies/μl and 70 copies/μl. Coefficient of variance (CV) was respectively 1.27%, 0.20%, 0.82%; 2.12%, 1.74%, and 1.40%. EKV-2 gene was detected in both whole blood and serum of a returnee from Angola.@*Conclusions@#The first EKV-2 gene was confirmed in both whole blood and serum of a returnee from Angola by real-time RT-PCR..

Factors affecting the fit of respirators in the biosafety level 3 laboratory / 中华实验和临床病毒学杂志

Objective@#To test the performance of two kinds of protective respirators (3M9332, 3M1860) used in Biosafety Level 3 Laboratory (BSL-3) to ensure whether it can fit well in different populations in order to reduce the risk of infection in the process of operating high-risk pathogenic microorganisms.@*Methods@#Using a 8038PortaCount®Pro+ respirator fit tester to examine the fitness of personnel wear 3M 1860 N95 and 9332 N99 respirators. The influence factors such as gender, age, shape of respirator were evaluated. Statistical analysis was performed by the fit factor(FF) of assigned actions.@*Results@#A total of 62 people conducted the respirator fit test, of which 45 people passed the test of 3M9332 respirators, the pass rate was 72.58%; only 6 people passed the test of 3M1860 respirators, the pass rate was 9.67%. The pass rate of two different types of respirators was analyzed statistically, P=0.000. The P value of assigned actions in deep breathing, head side to side, head up and down, talking, bending over were respectively 0.094, 0.076, 0.542, 0.000, 0.000.@*Conclusions@#The experimental results showed that the type of respirator had a significant effect on respirator fit in this study, but gender and age had no significant effect on factors. As to reduce the risk of leakage of the respirator, redundant actions should try to be avoided, especially loud talking and bending in the course of the experiment.

Establishment of Quality Control System of Nucleic Acid Detection for Ebola Virus in Sierra Leone-China Friendship Biological Safety Laboratory / 病毒学报

The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.

Establishment of an inactivated viral particle-based ELISA for the detection of anti-MERS-CoV anti-bodies / 中华微生物学和免疫学杂志

Objective To establish an inactivated viral particle-based ELISA for the detection of antibodies against Middle East respiratory syndrome coronavirus (MERS-CoV) in serum samples collected from a MERS-CoV associated case. Methods Serum samples were collected from 10 newborns and 40 healthy adults. A viral particle-based ELISA was established by using the inactivated MERS-CoV virions as antigen. The levels of IgM and IgG antibodies in the serum samples were detected by the established ELISA and the cut-off values for positive detection were determined. Then the inactivated MERS-CoV virion-based ELISA was used to detect the antibodies against MERS-CoV in 5 serum samples collected from the first im-ported MERS case in China. Results The cut-off values of IgM and IgG antibodies in serum samples for ELISA were determined to be A450 readings of 0. 32 and 0. 42, respectively. The titers of IgM and IgG anti-bodies in serum samples collected at early admission to hospital from the first imported MERS case in China were both 1 ︰ 40. Seroconversion occurred 2 weeks after his admission to hospital with the titers of IgM and IgG reaching to 1 ︰ 320. Conclusion The inactivated MERS-CoV virion-based ELISA was established successfully and could be used for the detection of serum antibodies (IgG and IgM) in MERS associated cases.

The expression of Th1/Th2 type cytokines and clinical significance in Uyghur cervical cancer / 实用肿瘤学杂志

Objective To assess the immune level of cervical cancer patients and precancerous lesions patients,and investigate the association of Th 1 /Th2 type cytokines expression and clinical pathologic factors in the plasma of Uyghur cervical cancer patients .Methods we collected peripheral blood specimens from cervical cancer patients,precancerous lesion(CINⅢ)patients and controls.The expressions of Th1 type cytokines IL-2, IFN-γand Th2 type cytokines IL-4,IL-10 in plasma were detected by ELISA .Receiver operating characteris-tic curve( ROC curve) was used to analyze the cytokines value of auxiliary diagnosis in cervical cancer .Results The results showed that compared with control group ,the expressions of Th1 type cytokines IL-2,IFN-γin cer-vical cancer and precancerous lesion group were significantly reduced (P<0.05).The expressions of Th2 type cytokines IL -4, IL -10 in cervical cancer and precancerous lesion group were significantly increased ( P <0.05).in cervical cancer group,IL -10 expression gradually increased with tumor pathological staging (P <0.05).IL-2 expression level in Uyghur cervical cancer patients was significantly lower than in Han patients (P<0.05).Compared cervical cancer group with control group ,AUC of IL-2,IFN-γ,IL-4 and IL-10 were 0.979,0.766,0.736 and 0.903.Conclusion Cellular immune level of cervical cancer patients was low and Th1/Th2 shift has occurred , which suggests that it may be one of the mechanisms in immune escape of tumor cells.Th1 /Th2 type cytokines detection has a certain significance for auxiliary diagnosis of cervical cancer .Be-sides,the decrease of IL-2 expression may play an important role in the occurrence and development of Uyghur patients with cervical cancer .