Journal of virological methods最新文献

{"title":"Development of neutralization tests using single-round infectious particles and cytopathic effect as an alternative method for measuring antibody titers against Japanese encephalitis virus in national epidemiological surveillance program of vaccine-preventable diseases in Japan","authors":"Shunsuke Yazawa ,&nbsp;Yumiko Saga ,&nbsp;Mami Matsuda ,&nbsp;Ryosuke Suzuki ,&nbsp;Shigeru Tajima ,&nbsp;Chang-Kweng Lim ,&nbsp;Hideki Tani","doi":"10.1016/j.jviromet.2025.115163","DOIUrl":"10.1016/j.jviromet.2025.115163","url":null,"abstract":"<div><div>The “National Epidemiological Surveillance of Vaccine-Preventable Diseases” is conducted annually in Japan for various infectious diseases. A susceptibility survey for Japanese encephalitis was conducted by measuring antibody titers using a focus-forming assay. However, this method is complicated to operate and hard to count small focuses; therefore, the development of new measurement methods is required. In this study, we developed and evaluated a single-round infectious particles (SRIPs) assay method and a cytopathic effect (CPE) assay method. The SRIPs assay showed a strong correlation (R = 0.94) with the focus-forming assay. The SRIPs assay method is not only simple and can be expected to improve accuracy, but also has the advantages of extremely low pathogenicity and safety for the operator. The CPE assay also showed a strong correlation (R = 0.80) with the focus-forming assay. It has the advantage of requiring almost no reagents and is easy to interpret. Both SRIPs and CPE assay methods can be used as alternatives to the focus-forming assay method, but further characterization of each method is necessary before one is selected for routine use.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"336 ","pages":"Article 115163"},"PeriodicalIF":2.2,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143807649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of bioinformatic pipelines for virus monitoring using nanopore sequence data: a systematic assessment.","authors":"Nick Laurenz Kaiser, Martin H Groschup, Balal Sadeghi","doi":"10.1016/j.jviromet.2025.115153","DOIUrl":"https://doi.org/10.1016/j.jviromet.2025.115153","url":null,"abstract":"<p><p>Nanopore sequencing has proven to be a promising technique in virus surveillance efforts, especially due to the portability of its sequencers. In order to process the long, error-prone reads generated, specialised bioinformatic programs are required. These can be run automatically within pipelines so as to effectively provide decisionmakers with all relevant information about the molecular characteristics of a virus. The purpose of this systematic assessment was to identify pipelines that are suitable for virus surveillance programs using nanopore sequencing. Promising candidates were then compared in terms of their functional scope. Of 239 initial papers, 22 pipelines were tested, of which six were included in the final assessment. The four pipelines that were exclusively available offline were each missing individual downstream analysis steps considered in our assessment. The other two executed all steps. One of these was only available online and subject to a charge, while the other was freely available both online and offline. While we were able to identify two pipelines that are broadly suitable for virus surveillance using nanopore sequencing, we discovered two major shortcomings in this domain. None of the pipelines integrated basecalling, the initial step of data processing. In addition, there was no pipeline that was easy to install and provided all relevant analysis results with a single program call. We therefore see a need for the development of a pipeline that incorporates both aspects.</p>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":" ","pages":"115153"},"PeriodicalIF":2.2,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an allele-specific quantitative polymerase chain reaction assay for differentiating the RLB 106 strain from the wild-type viruses of Varicellovirus bovinealpha1","authors":"Keigo Ikeda ,&nbsp;Yuto Suda ,&nbsp;Hisayuki Tomochi ,&nbsp;Shinichi Hatama ,&nbsp;Yoshifumi Iwamaru","doi":"10.1016/j.jviromet.2025.115148","DOIUrl":"10.1016/j.jviromet.2025.115148","url":null,"abstract":"<div><div><em>Varicellovirus bovinealpha1</em> (BoAHV1) is an important causal agent of various pathological conditions, such as respiratory and reproductive diseases, in cattle. An intranasal vaccine containing a temperature-sensitive mutant RLB 106 strain is used to control BoAHV1-related diseases. To monitor the invasion of BoAHV1 wild-type viruses in vaccinated populations, it is essential to develop a simple method for differentiating between the RLB 106 strain and BoAHV1 wild-type viruses. In this study, we developed an allele-specific quantitative polymerase chain reaction (AS-qPCR) assay that targets the point mutation G23136A located in UL40 for detecting the RLB 106 strain. We calculated the difference in Cq values (ΔCq) obtained from a paired qPCR using each point mutation site-targeting primer set and established the cutoff ΔCq for differentiation. The accuracy of differentiation was confirmed by the DNA partial sequencing of UL40. Results demonstrated that differentiation by AS-qPCR was consistent with the results obtained by DNA sequencing, and the field isolates classified as the RLB 106 strain exhibited a temperature-sensitive phenotype. Hence, the AS-qPCR assay developed in this study could be a promising method for the simple differentiation between the RLB 106 strain and wild-type viruses in a single-step reaction.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"336 ","pages":"Article 115148"},"PeriodicalIF":2.2,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of EBOV GP IgG antibody reactivity: Results from two immunoassays in the Democratic Republic of the Congo","authors":"Sydney Merritt ,&nbsp;Megan Halbrook ,&nbsp;Jean Paul Kompany ,&nbsp;Prabha Chandrasekaran ,&nbsp;Olivia A. Smith ,&nbsp;Nicole A. Hoff ,&nbsp;Merly Tambu ,&nbsp;Skylar A. Martin ,&nbsp;Teri Ann Wong ,&nbsp;Amie Jarra ,&nbsp;Angelica L. Barrall ,&nbsp;Kamy Musene ,&nbsp;Michael Beya ,&nbsp;Robert Orr ,&nbsp;Todd Myers ,&nbsp;Tracy MacGill ,&nbsp;Lisa E. Hensley ,&nbsp;Jean-Jacques Muyembe-Tamfum ,&nbsp;Didine Kaba ,&nbsp;Irina Maljkovic Berry ,&nbsp;Anne W. Rimoin","doi":"10.1016/j.jviromet.2025.115154","DOIUrl":"10.1016/j.jviromet.2025.115154","url":null,"abstract":"<div><div>Ebola virus (EBOV) is a highly infectious pathogen, and its long-term consequences continue to be investigated. With its high fatality rate and potential for reinfection or latent infection, continued development of research tools is of utmost importance. Using a cohort (n = 503) of existing bio-banked specimens from the Democratic Republic of the Congo (DRC) two EBOV glycoprotein (GP) immunoglobulin G (IgG) antibody-detection assays were compared: the gold-standard Filovirus Animal Non-Clinical Group (FANG) and a Multiplex bead-based Immunoassay (MIA) with seven pan-filoviral targets. As not all immunoassays have been shown to detect a vaccine-induced immune response, and previous EBOV serosurveillance has been primarily conducted with singleplex technology, this MIA was assessed as an additional resource. Among the cohort, as sample seroreactivity increased, assay correlation increased (r²=0.80). Correlation was sustained among sub-populations of the cohort—in detecting natural immunity among survivors and vaccine-derived responses. Additionally, when results were binarized by seroreactivity, there was high correlation between the two assays (kappa=0.70) with 71 serodiscordant samples. These data indicate that the MIA is an apt alternative to the singleplex FANG assay in detecting relative seroreactivity and can be used as a potential tool for widespread pan-filovirus serosurveillance in the DRC and similar contexts.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"336 ","pages":"Article 115154"},"PeriodicalIF":2.2,"publicationDate":"2025-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of an infectious subgenomic amplicons reverse genetics protocol for the rescue of synthetic coronaviruses","authors":"Ilaria Puglia ,&nbsp;Marialuigia Caporale ,&nbsp;Giovanni Di Teodoro ,&nbsp;Massimo Spedicato ,&nbsp;Francesca Profeta ,&nbsp;Maurilia Marcacci ,&nbsp;Chiara Di Pancrazio ,&nbsp;Fabrizia Valleriani ,&nbsp;Emanuela Rossi ,&nbsp;Heidi Auerswald ,&nbsp;Alessio Lorusso","doi":"10.1016/j.jviromet.2025.115152","DOIUrl":"10.1016/j.jviromet.2025.115152","url":null,"abstract":"<div><div>Reverse genetics (rg) systems are indispensable tools for investigating the pathogenesis of RNA viruses, facilitating vaccine design, and advancing antiviral therapeutic strategies. In this study, we optimized the Infectious Subgenomic Amplicons (ISA) method for generating synthetic r-wt SARS-CoV-2 Wuhan-Hu-1. This system was validated by demonstrating the successful rescue of infectious viral particles from overlapping DNA fragments and their propagation <em>in vitro</em>. Sequencing confirmed 100 % identity of the recovered virus with the Wuhan-Hu-1 reference genome. Importantly, <em>in vivo</em> experiments using K18-hACE2 mice revealed that the r-wt SARS-CoV-2 Wuhan-Hu-1 strain caused clinical symptoms, weight loss, and mortality comparable to those induced by a virulent SARS-CoV-2 field variant. This ISA rg method offers a rapid and reproducible approach to generating synthetic coronaviruses, with potential applications in pathogenesis studies, antiviral testing, and vaccine development.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"336 ","pages":"Article 115152"},"PeriodicalIF":2.2,"publicationDate":"2025-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143795729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The potential application of digital PCR in detecting different SARS-CoV-2 viral loads","authors":"Chunyan Chang ,&nbsp;Lingling Li ,&nbsp;Yating Guo ,&nbsp;Li Ji ,&nbsp;Jinyue Tian ,&nbsp;Shenglin Xu ,&nbsp;Xiuhong Zhang ,&nbsp;Xinyi Jiang ,&nbsp;Weizhen Qiao","doi":"10.1016/j.jviromet.2025.115151","DOIUrl":"10.1016/j.jviromet.2025.115151","url":null,"abstract":"<div><div>Rapid, effective, and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial. It is essential to control the spread of the virus and ensure accurate treatment for the disease. In the study, a total of 170 clinical specimens from 164 patients were collected and analyzed through digital PCR (dPCR) and real time quantitative reverse transcription PCR (RT-qPCR). The results showed an 86.41 % agreement between dPCR and RT-qPCR, with differences primarily noted in suspected cases. RT-qPCR exhibited a sensitivity of 84.78 %, specificity of 95.83 %, and accuracy of 86.42 %, which were comparatively lower than the 100 % accuracy of dPCR. Subsequently, we explored the potential correlation between these two methodologies based on Ct value groups. A strong negative correlation was observed between RT-qPCR and dPCR techniques in the Ct value group between 25 and 35, while the correlation was weakest in the Ct &gt; 35 group. Moreover, the concordance rate for detecting the ORF1 (142/162) gene by RT-qPCR was lower compared to that of the N gene (149/162). Additionally, nucleic acid concentrations for ORF1 gene detection were lower than those for N gene detection in dPCR. In conclusion, this study shows that dPCR provides more reliable detection than RT-qPCR, especially for samples with low viral loads. Furthermore, dPCR effectively tracked changes in viral load during hospitalization, facilitating the diagnosis and treatment of COVID-19.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"335 ","pages":"Article 115151"},"PeriodicalIF":2.2,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143768642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genotypic analysis of rhinovirus and human respiratory syncytial virus in sudden unexpected death in infancy cases at Tygerberg Hospital, Cape Town, South Africa","authors":"Corena de Beer,&nbsp;Hameer Deepak Vanmali","doi":"10.1016/j.jviromet.2025.115150","DOIUrl":"10.1016/j.jviromet.2025.115150","url":null,"abstract":"<div><div>Infant mortality remains a major global concern. Sudden unexpected death in infancy (SUDI) is reported globally and an infant mortality rate of 23.129 per 1 000 live births has been reported in the Western Cape, South Africa, in 2024. Infections are often confirmed in SUDI cases admitted to the Tygerberg Medico-legal Mortuary in Cape Town, but molecular diversity in respiratory viruses is underreported. A total of 162 previously confirmed polymerase chain reaction (PCR)-positive trachea and / or lung samples from SUDI cases collected between 2015 and 2019 were retested for either rhinovirus or human respiratory syncytial virus (RSV). Sixty-four samples were positive for rhinovirus and 15 for RSV. Results from 5 of all positive samples were outside the PCR assay amplification limits determined by the cycle threshold (Ct) value and were excluded. Another 4 samples did not amplify, and the remaining 70 underwent subsequent sequencing, but successful sequences could only be obtained in 53 samples. All three rhinovirus (A, B and C) genotypes were identified, with RV-A most prevalent, followed by RV-C and RV-B. RSV-A and RSV-B were detected equally, and after amino acid alignment, 20 amino acid duplication and nine substitutions were found that confirmed two RSV-BA9 genotypes. This study describes the molecular and phylogenetic characterisation of specific respiratory viruses in SUDI cases in South Africa. However, the rapid decline in viral viability in post-mortem samples does not allow correlation between viral genotypes and cause of death or disease severity. Future prospective studies should therefore investigate temporality and associations between specific viral strains and clinical disease severity and mortality.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"335 ","pages":"Article 115150"},"PeriodicalIF":2.2,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143715265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rethinking statistical approaches for serological data analysis for viral surveillance","authors":"Morgan P. Kain ,&nbsp;Jonathan H. Epstein ,&nbsp;Noam Ross","doi":"10.1016/j.jviromet.2025.115149","DOIUrl":"10.1016/j.jviromet.2025.115149","url":null,"abstract":"<div><div>A robust serological surveillance system for zoonotic pathogens is imperative for both early detection and advancing knowledge of emerging diseases. A statistical analysis plan that is aligned to research and epidemiological goals requires a purposeful choice among alternative methods for differentiating seronegative from seropositive samples, estimating seroprevalence, and estimating risk factors associated with seropositivity. The common standard deviation-based cutoff (e.g., 3sd) approach is simple to implement and understand, but fails to appropriately propagate uncertainty in serostatus assignments to any risk factor analysis. Methods such as Gaussian mixture models, which jointly estimate serostatus, risk factors, and their uncertainty, can alleviate the dichotomy created by the cutoff approach. Yet, because of a lack of empirical guidance of method performance, it remains difficult to choose a robust analysis method for a given serological dataset. Here we examine the performance of both cutoff and clustering approaches using simulated datasets that represent the epidemiological, biological, and immunological data generation process. We focus on understudied pathogens for which validated serological assays do not exist, as is common in emerging viruses in wildlife. We quantify coverage (the proportion of time 95 % confidence intervals contain the true value) and bias (systematic differences between true values and model point estimates) of model estimates for individual serostatus assignments, population seroprevalence, and regression coefficients for serostatus risk factors. In nearly all scenarios, Bayesian mixture models provide the highest coverage and lowest bias. Only with very low seroprevalence (∼ &lt; 3 %) and large differences in signal between seronegative and seropositive individuals will a cutoff provide low bias and near-nominal coverage. Given poor coverage of risk factor regression coefficients, we advise against using a cutoff approach for quantifying determinants of seropositivity.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"335 ","pages":"Article 115149"},"PeriodicalIF":2.2,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143692628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The stability and elimination of mammalian enveloped and non-enveloped respiratory and enteric viruses in indoor air: Testing using a room-sized aerobiology chamber","authors":"Bahram Zargar ,&nbsp;Syed A. Sattar ,&nbsp;Julie McKinney ,&nbsp;M. Khalid Ijaz","doi":"10.1016/j.jviromet.2025.115144","DOIUrl":"10.1016/j.jviromet.2025.115144","url":null,"abstract":"<div><div>We assessed the viability of aerosolized human betacoronavirus OC43 (HCoV-OC43; ATCC VR-1558), human rhinovirus-14 (RV-14; ATCC VR-284) and feline calicivirus (FCV; ATCC VR-782) as representative enveloped and non-enveloped respiratory and enteric viruses of mammals in indoor air under ambient conditions (relative humidity 50 ± 10 % and air temperature 22 ± 2°C) using a room-sized (25 m³; 900 ft³) aerobiology chamber. All virus suspensions contained a soil load to simulate the presence of body fluids and they were separately aerosolized into the chamber using a six-jet Collison nebulizer. A muffin fan was used to uniformly mix the air inside the chamber and to keep the aerosols airborne. A slit sampler with Petri plates containing 3 % (wt./vol) gelatin was used to collect the air samples. The gelatin was liquefied in an incubator and assayed for infectious virus as plaque-forming units (PFU). The rates of biological decay of HCoV-OC43, RV-14 and FCV were 0.0052 ± 0.00026, 0.0034 ± 0.0027 and 0.0081 ± 0.0031 (as log₁₀ PFU/m³/min), respectively. We also assessed a HEPA filter-based stand-alone air purifier against the experimentally aerosolized viruses and the device could demonstrate <em>&gt;</em> 3-log₁₀ reductions in the viability of the three viruses in 46, 62 and 41 minutes, respectively. Therefore, we can now investigate the stability of mammalian viruses in indoor air as well as air decontamination technologies against them under field-relevant conditions.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"335 ","pages":"Article 115144"},"PeriodicalIF":2.2,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143597342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Complete inactivation of orthoflavi- and alphaviruses by acetone for safe titering by ELISA","authors":"Robson K. Loterio ,&nbsp;Katherine Rosevear ,&nbsp;Kathryn Edenborough ,&nbsp;Johanna E. Fraser","doi":"10.1016/j.jviromet.2025.115146","DOIUrl":"10.1016/j.jviromet.2025.115146","url":null,"abstract":"<div><div>The tissue culture infectious dose 50 (TCID₅₀) end-point dilution assay is the gold-standard assay to titer viruses with negligible or ambiguous cytopathic effects. The assay’s specificity is improved when followed by an Enzyme-Linked Immunosorbent Assay (ELISA) to detect viral antigens. Cells infected with mosquito-borne orthoflavi- and alphaviruses are fixed after TCID₅₀, prior to ELISA, using paraformaldehyde (PFA) or acetone. While 4 % PFA has been shown to effectively inactivate these viruses for safe handling in low biocontainment conditions, equivalent studies have not been reported for standard acetone fixation methods (20 % acetone for 24 hours at 4°C). This study evaluated the inactivation efficacy of acetone on orthoflavi- and alphaviruses using dengue virus (DENV) and Ross River virus (RRV), as exemplar viruses from each genus, respectively. We show that 50 % acetone and 4 % PFA fully inactivate DENV and RRV, but 20 % acetone does not reduce the infectivity of these viruses. Importantly, ELISA-based detection of DENV- and RRV-infected cells fixed with 50 % acetone was effective, with calculated titres comparable to cells treated with 20 % acetone. Together, our results inform a fixation method for titrating orthoflavi- and alphavirus samples by TCID₅₀/ELISA, ensuring the safe handling and processing of these viruses under low biocontainment conditions.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"335 ","pages":"Article 115146"},"PeriodicalIF":2.2,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143577045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}