Journal of virological methods最新文献

{"title":"Real-world performance of the NeuMoDx™ HCV Quant Test for quantification of hepatitis C virus (HCV)-RNA","authors":"Nadine Lübke ,&nbsp;Andreas Walker ,&nbsp;Martin Obermeier ,&nbsp;Jennifer Camdereli ,&nbsp;Martha Paluschinski ,&nbsp;Lara Walotka ,&nbsp;Anna-Kathrin Schupp ,&nbsp;Inga Tometten ,&nbsp;Sandra Hauka ,&nbsp;Eva Heger ,&nbsp;Jörg Timm","doi":"10.1016/j.jviromet.2024.114937","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114937","url":null,"abstract":"<div><p>Quantification of hepatitis C virus (HCV)-RNA in serum or plasma samples is an essential parameter in HCV diagnostics. Here, the NeuMoDx™Molecular System (Qiagen) was tested for the most common HCV genotypes and compared to the cobas c6800 system (Roche).</p><p>HCV-RNA from 131 plasma/serum samples from chronically infected patients was determined in parallel on the NeuMoDx and c6800 systems. Linearity was analysed using the four most common HCV genotypes (1−4) in our cohort. The coefficient of variation (CV) within (intra-assay) and between (inter-assay) runs was calculated based on HCV-RNA concentration. Quantitative HCV-RNA results were highly correlated on both test systems (R² = 0.7947; y = 0.94 x + 0.37). On average, the NeuMoDx and c6800 HCV RNA levels showed a mean difference of only 0.05 log10 IU/mL but with a broad distribution (±1.2 2 x SD). The NeuMoDx demonstrated very good linearity across all HCV genotypes tested at concentrations between 1.7 and 6.2 log10 IU/mL (R² range: 0.9257–0.9991) with the highest mean coefficient of determination for genotype 1 (R² = 0.9909). The mean intra- and inter-assay CV for both serum and plasma samples was &lt;5 %. The NeuMoDx HCV-RNA Assay demonstrates high subtype-independent comparability, linearity, and reproducibility for the quantification of HCV-RNA in serum and plasma samples from chronically infected patients.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0166093424000612/pdfft?md5=d68fece79f91e9f69855b776b50b5cb7&pid=1-s2.0-S0166093424000612-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140632599","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":"A validated in-house assay for HIV drug resistance mutation surveillance from dried blood spot specimens","authors":"Bronwyn Neufeld ,&nbsp;Chantal Munyuza ,&nbsp;Alexandria Reimer ,&nbsp;Rupert Capiña ,&nbsp;Emma R. Lee ,&nbsp;Marissa Becker ,&nbsp;Paul Sandstrom ,&nbsp;Hezhao Ji ,&nbsp;François Cholette","doi":"10.1016/j.jviromet.2024.114939","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114939","url":null,"abstract":"<div><p>Despite increasing scale-up of antiretroviral therapy (ART) coverage, challenges related to adherence and HIV drug resistance (HIVDR) remain. The high cost of HIVDR surveillance is a persistent challenge with implementation in resource-constrained settings. Dried blood spot (DBS) specimens have been demonstrated to be a feasible alternative to plasma or serum for HIVDR genotyping and are more suitable for lower resource settings. There is a need for affordable HIVDR genotyping assays which can amplify HIV-1 sequences from DBS specimens, particularly those with low viral loads, at a low cost. Here, we present an in-house assay capable of reliably amplifying HIV-1 protease and partial reverse transcriptase genes from DBS specimens, which covers the complete World Health Organization 2009 list of drug resistance mutations under surveillance. DBS specimens were prepared using whole blood spiked with HIV-1 at concentrations of 10,000, 5000, 1000, and 500 copies/mL (n=30 for each concentration). Specimens were tested in triplicate. A two-step approach was used consisting of cDNA synthesis followed by nested PCR. The limit of detection of the assay was calculated to be approximately 5000 (95<!--> <!-->% CI: 3200–10,700) copies/mL for the protease gene and 3600 (95<!--> <!-->% CI: 2200–10,000) copies/mL for reverse transcriptase. The assay was observed to be most sensitive with higher viral load specimens (97.8<!--> <!-->% [95<!--> <!-->% CI: 92.2–99.7]) for both protease and reverse transcriptase at 10,000 copies/mL with performance decreasing with the use of specimens with lower viral loads (46.7<!--> <!-->% [36.1–57.5] and 60.0<!--> <!-->% [49.1–70.2] at 500 copies/mL for protease and reverse transcriptase, respectively). Ultimately, this assay presents a promising opportunity for use in resource-constrained settings. Future work should involve validation under field conditions including sub-optimal storage conditions and preparation of DBS with fingerprick blood in order to accurately reflect real-world collection scenarios.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0166093424000636/pdfft?md5=33d7e82bc47839f120e0a31109768c20&pid=1-s2.0-S0166093424000636-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140559157","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":"Clinical validation of SARS-CoV-2 electrochemical immunosensor based on the spike-ACE2 complex","authors":"Viviana Vásquez,&nbsp;Jahir Orozco","doi":"10.1016/j.jviromet.2024.114940","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114940","url":null,"abstract":"<div><h3>Background and aims</h3><p>Advances in health, especially in prevention, diagnosis, and treatment, have significantly impacted the way of facing emerging infectious diseases. Yet, events such as the COVID-19 pandemic have shown that there is still a long way to go. Therefore, an urgent need exists for portable and easily deployable point-of-care (POC) detection tools. Biosensors at the POC remain in the laboratory in an analytical characterization step and are not yet mature enough to reach the market massively. In this context, it is necessary to progress in validating these devices to demonstrate their relevance in detecting different disease biomarkers. This work reports on the clinical validation of an electrochemical immunosensor for detecting SARS-CoV-2.</p></div><div><h3>Methods</h3><p>A monocentric retrospective cohort study was conducted with 150 random nasopharyngeal swabs or tracheal aspiration samples tested by RT-PCR. The immunosensor based on magnetic beads and chronoamperometry detected SARS-CoV-2 through the spike-angiotensin-converting protein (ACE2) immunocomplex.</p></div><div><h3>Results</h3><p>This biosensor demonstrated 96.04 % clinical sensitivity and 87.75 % clinical specificity in detecting SARS-CoV-2 in the samples, highly correlated with the RT-PCR gold standard.</p></div><div><h3>Conclusions</h3><p>It demonstrates the potential of electrochemical biosensors to be implemented as highly sensitive and easily deployable detection strategies even in remote locations.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140632642","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 and analytical validation of a novel quantitative PCR assay for the detection of Trachemys herpesvirus 1","authors":"Kaitlin A. Moorhead ,&nbsp;Laura A. Adamovicz ,&nbsp;Matthew C. Allender","doi":"10.1016/j.jviromet.2024.114941","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114941","url":null,"abstract":"<div><p>Emerging infectious diseases are a threat that contributes to the decline of global chelonian species. Herpesviruses are among the most impactful pathogens described in chelonians and are frequently associated with a range of presentations across hosts with the potential for severe morbidity and mortality. <em>Trachemys</em> herpesvirus 1 (TrHV1) has been reported in red-eared and yellow-bellied sliders (<em>Trachemys scripta elegans</em> and <em>Trachemys scripta scripta</em>, respectively) but is largely understudied. Invasive red-eared sliders may serve as a reservoir for transmission to sympatric native species. This study aimed to develop a sensitive and specific quantitative real-time PCR (qPCR) assay for the detection of TrHV1 DNA to aid in the characterization of the epidemiology of this virus in aquatic turtles. Two TaqMan-MGB FAM-dye labeled primer-probe sets were designed and evaluated using plasmid dilutions. The higher performing assay was specific for TrHV1 DNA and had a linear dynamic range of 1.0 × 10⁷ to 1.0 × 10¹ copies per reaction with an R² of 0.999, slope of −3.386, and efficiency of 97.39%. The limit of detection was 10¹ copies per reaction, and there was no loss of reaction efficiency in the presence of TrHV1-negative chelonian oral-cloacal DNA. Overall, the <em>Trachemys</em> herpesvirus 1 assay meets established criteria for acceptable qPCR assays and will be a valuable tool in characterizing the epidemiology of <em>Trachemys</em> herpesvirus 1 in chelonians.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S016609342400065X/pdfft?md5=864e936309877f0e837a5fde44deb8aa&pid=1-s2.0-S016609342400065X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140607360","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":"RSV-GenoScan: An automated pipeline for whole-genome human respiratory syncytial virus (RSV) sequence analysis","authors":"Alexandre Dosbaa ,&nbsp;Romane Guilbaud ,&nbsp;Anna-Maria Franco Yusti ,&nbsp;Valentine Marie Ferré ,&nbsp;Charlotte Charpentier ,&nbsp;Diane Descamps ,&nbsp;Quentin Le Hingrat ,&nbsp;Romain Coppée","doi":"10.1016/j.jviromet.2024.114938","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114938","url":null,"abstract":"<div><h3>Background</h3><p>Advances in high-throughput sequencing (HTS) technologies and reductions in sequencing costs have revolutionised the study of genomics and molecular biology by making whole-genome sequencing (WGS) accessible to many laboratories. However, the analysis of WGS data requires significant computational effort, which is the major drawback in implementing WGS as a routine laboratory technique.</p></div><div><h3>Objective</h3><p>Automated pipelines have been developed to overcome this issue, but they do not exist for all organisms. This is the case for human respiratory syncytial virus (RSV), which is a leading cause of lower respiratory tract infections in infants, the elderly, and immunocompromised adults.</p></div><div><h3>Results</h3><p>We present RSV-GenoScan, a fast and easy-to-use pipeline for WGS analysis of RSV generated by HTS on Illumina or Nanopore platforms. RSV-GenoScan automates the WGS analysis steps directly from the raw sequence data. The pipeline filters the sequence data, maps the reads to the RSV reference genomes, generates a consensus sequence, identifies the RSV subgroup, and lists amino acid mutations, insertions and deletions in the F and G viral genes. This enables the rapid identification of mutations in these coding genes that are known to confer resistance to monoclonal antibodies.</p></div><div><h3>Availability</h3><p>RSV-GenoScan is freely available at <span>https://github.com/AlexandreD-bio/RSV-GenoScan</span><svg><path></path></svg>.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0166093424000624/pdfft?md5=f24cdd06dbedfd40bbdb57ad56f718a0&pid=1-s2.0-S0166093424000624-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140554636","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":"Insertion of foreign genes into the simian varicella virus genome by Tn7-mediated site-specific transposition","authors":"Wayne L. Gray","doi":"10.1016/j.jviromet.2024.114936","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114936","url":null,"abstract":"<div><p>A Tn7-transposition approach was utilized for site-specific insertion of foreign genes into the genome of simian varicella virus (SVV), the causative agent of simian varicella in nonhuman primates. The severe acute respiratory syndrome coronavirus (SARS-CoV-2) nucleocapsid (N) gene and receptor binding domain (RBD) of the spike gene were inserted into the ORF 14 region of the SVV genome cloned into a bacterial artificial chromosome and then transfected into Vero cells to generate the infectious recombinant SVV (rSVV). The rSVV replicated efficiently in infected Vero cells and expressed the N and RBD antigens as indicated by immunoblot and immunofluorescence assays. Tn7-mediated transposition provides a rapid and efficient method for constructing rSVVs which may be evaluated as live-attenuated vaccines.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140559156","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":"Construction and characterization of recombinant senecavirus A expressing secreted luciferase for antiviral screening","authors":"Hao Wang ,&nbsp;Yongfang Mo ,&nbsp;Wenbo Liu ,&nbsp;Qijie He ,&nbsp;Tongwei Ren ,&nbsp;Kang Ouyang ,&nbsp;Ying Chen ,&nbsp;Weijian Huang ,&nbsp;Zuzhang Wei","doi":"10.1016/j.jviromet.2024.114932","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114932","url":null,"abstract":"<div><p>Senecavirus A (SVA) is a newly identified picornavirus associated with swine vesicular disease and neonatal mortality. The development of an SVA incorporating an exogenous reporter gene provides a powerful tool for viral research. In this study, we successfully constructed a recombinant SVA expressing Gaussia Luciferase (Gluc), termed rSVA-Gluc. The growth kinetics of rSVA-Gluc in BHK-21 cells were found to be comparable to those of the parental virus, and Gluc activity paralleled the virus growth curve. Genetic analysis revealed stable inheritance of the inserted reporter protein genes for at least six generations. We evaluated the utility of rSVA-Gluc in antiviral drug screening, and the results highlighted its potential as an effective tool for such purposes against SVA.</p></div><div><h3>Data availability statement</h3><p>The data that support the findings of this study are available on request from the corresponding author.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140350000","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":"Intracellular localized heterogeneous protein franking by a transmembrane domain of GP64 is sufficient to be assembled on budded virions of Bombyx mori nucleopolyhedrovirus","authors":"Yufeng Hao ,&nbsp;Na Liu ,&nbsp;Jingfeng Li ,&nbsp;Stephen Baffour Gyawu ,&nbsp;Ogone Emeldah Setshogo ,&nbsp;Jinshan Huang ,&nbsp;Bifang Hao","doi":"10.1016/j.jviromet.2024.114933","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114933","url":null,"abstract":"<div><p>Baculovirus has been widely used for foreign protein expression in biomedical studies, and budded virus (BV) surface display has developed into an important research tool for heterogenous membrane protein studies. The basic strategy of surface display is to construct a recombinant virus where the target gene is fused with a complete or partial <em>gp64</em> gene. In this study, we further investigate and develop this BV surface displaying strategy. We constructed stable insect cell lines to express the target protein flanking with different regions of signal peptide (SP) and GP64 transmembrane domain (TMD). Subsequently, recombinant BmNPV was used to infect the cell, and the integration of heterogeneous protein into BV was detected. The results indicated that deletion of the n-region of SP (SP^Δn) decreased the incorporation rate more than that of the full-length SP. However, the incorporation rate of the protein fused with h and c-region deletion of SP (SP^Δh-c) was significantly enhanced by 35–40 times compare to full-length SP. Moreover, the foreign protein without SP and TMD failed to display on the BV, while the integration of foreign proteins with GP64 TMD fusion at the c-terminal was significantly enhanced by 12–26 times compared to the control. Thus, these new strategies developed the BV surface display system further.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140549715","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":"Evaluation of an automated platform for the detection of HEV RNA in plasma and stool","authors":"Pauline Sottil ,&nbsp;Sébastien Lhomme ,&nbsp;Karine Saune ,&nbsp;Soheil El Hayani ,&nbsp;Kévin Oliveira-Mendes ,&nbsp;Jean-Marie Peron ,&nbsp;Nassim Kamar ,&nbsp;Jacques Izopet ,&nbsp;Florence Abravanel","doi":"10.1016/j.jviromet.2024.114920","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114920","url":null,"abstract":"<div><h3>Introduction</h3><p>We evaluated the performance of the automated Altostar HEV RNA platform for detecting HEV RNA.</p></div><div><h3>Methods and results</h3><p>Clinical performance was determined by testing 81 plasma samples and 10 fecal samples manually quantified previously with the Realstar RT-PCR assay using the Magnapure instrument for extraction. The assays were concordant for 79/81 plasma samples (97.5%) and 10/10 (100%) fecal samples. The two plasma samples that tested negative with the Altostar assay had a very low HEV RNA concentration (1.6 and 1.4 log₁₀ IU/ml). Quantitative results obtained with the automated platform and the manual workflow were highly correlated (ρ= 0.98, p&lt;0.01). The intra-run and inter-run standard deviation were 0.09 IU/ml and 0.13 IU/ml respectively. The assay was linear from 2 to 6 log IU/ml. The limit of detection determined by Probit analysis with the WHO HEV RNA standard was 7.6 [95% CI: 4.4–52.5] IU/ml.</p></div><div><h3>Conclusions</h3><p>The Altostar platform enables highly accurate testing for the detection of HEV RNA in stool and the quantification of HEV RNA in plasma. This allowed us to shorten turnaround times and to save time for the technical staff.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140543614","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":"Profiling of Groundnut bud necrosis orthotospovirus–responsive microRNA and their targets in tomato based on deep sequencing","authors":"M. Nivedha ,&nbsp;S. Harish ,&nbsp;K. Angappan ,&nbsp;G. Karthikeyan ,&nbsp;K.K. Kumar ,&nbsp;M. Murugan ,&nbsp;J. Infant Richard","doi":"10.1016/j.jviromet.2024.114924","DOIUrl":"https://doi.org/10.1016/j.jviromet.2024.114924","url":null,"abstract":"<div><p>Tomato, an extensively cultivated vegetable crop produces miRNAs in response to infection with <em>Groundnut bud necrosis orthotospovirus</em>, a viral pathogen causing significant economic losses. High-throughput miRNA sequencing was performed on tomato leaves inoculated with GBNV and mock-inoculated leaves as controls. Analysis revealed 73 known miRNAs belonging to 24 miRNA families, with variable expression levels. Interestingly, 39 miRNAs were upregulated, and 34 were downregulated in response to GBNV infection. Stem-loop quantitative reverse transcription PCR validated the differential expression of selected miRNAs. Additionally, 30 miRNA encoded proteins were identified to be involved in disease resistance and susceptibility. The miRNA-target interactions were found to play significant roles in cellular and metabolic activities, as well as modulating signaling pathways during the plant-virus interaction. The findings shed light on the intricate regulatory network of miRNAs in tomato response to viral infection and may contribute to developing strategies for improving crop protection against viral diseases.</p></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140618466","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}