Search Results (211)

Mycoplasma pneumoniae (MP) is a frequent cause of pediatric pneumonia, but the clinical relevance of respiratory coinfections remains incompletely defined. We conducted a retrospective cohort study of 192 hospitalized children with multiplex RT-PCR-confirmed MP infection between April 2023 and November 2025. Patients were classified as MP monoinfection (123/192, 64.1%), MP with viral coinfection (34/192, 17.7%), or MP with bacterial/mixed coinfection (35/192, 18.2%) and children with coinfection were younger than those with MP infection alone, with median ages of 6.5 years (IQR 1–11) in the viral coinfection group and 5 years (IQR 1–7) in the bacterial/mixed coinfection group, compared with 11 years (IQR 7–14) in children with MP alone (p < 0.001). Respiratory failure occurred in 36% of children with MP infection alone, 65% of those with viral coinfection, and 31% of those with bacterial/mixed coinfection. In adjusted logistic regression, viral coinfection was independently associated with higher odds of respiratory failure relative to MP infection alone (aOR 3.37, 95% CI 1.49–7.94; p = 0.004), whereas bacterial/mixed coinfection was not (aOR 0.85, 95% CI 0.35–2.03; p = 0.725). In multinomial regression, increasing age was inversely associated with both viral coinfection (RRR 0.86, 95% CI 0.79–0.94; p < 0.001) and bacterial/mixed coinfection (RRR 0.79, 95% CI 0.72–0.87; p < 0.001). In negative binomial regression, evidence for longer hospitalization in the viral coinfection group was weaker and imprecise (IRR 1.16, 95% CI 0.99–1.37; p = 0.069). Fibrinogen differed across etiologic groups, with the lowest values in bacterial/mixed coinfection (p = 0.004). In this hospitalized pediatric cohort, MP coinfection was more frequent in younger children, and viral co-detection showed the clearest association with respiratory failure. These findings should be interpreted in the context of the retrospective design, modest subgroup sizes, and the limitations of upper-airway multiplex PCR co-detection. Full article

Background: Following a significant decline during the 2020–2021 SARS-CoV-2 pandemic, Mycoplasma pneumoniae (MP) experienced a resurgence across Europe in 2023–2024. Although macrolide-resistant MP has increased globally, severe disease can occur even in the absence of resistance, which highlights the importance of rapid molecular characterization for clinical purposes. In this context, clinical severity is often improperly used as a surrogate marker of macrolide resistance, potentially driving unnecessary antibiotic escalation. Methods: We report a severe MP pneumonia occurring during the 2023–2024 resurgence and evaluate macrolide resistance through a rapid two-step workflow (Real Time-PCR screening for A2063G/A2064G followed by confirmatory 23S rRNA sequencing), to assess whether severity predicts resistance and to support antibiotic stewardship. Results: The patient developed acute hypoxic respiratory failure (PaO₂ 54.9 mmHg; P/F ratio 110), extensive centrilobular micronodules on chest CT imaging, significant systemic inflammation and elevated liver enzymes. Respiratory support was escalated from a Venturi mask to a high-flow nasal cannula and BiPAP. MP infection was confirmed by multiplex Real Time-PCR (RT-PCR) and supported by positive IgM/IgG serology. RT-PCR targeting A2063G/A2064G mutations revealed no resistance-associated variants, and Sanger sequencing of an 807 bp 23S rRNA fragment confirmed a wild-type genotype. Despite severe hypoxemic respiratory failure, no resistance-associated variants were detected, documenting a clear severity–genotype mismatch. Clinical and radiological improvement followed second-line antibiotic therapy. Conclusions: Severe MP pneumonia can occur despite the absence of macrolide resistance. During MP re-emergence, clinical severity should not be used to infer macrolide resistance. Integrating nucleic acid amplification test (NAAT) diagnosis with rapid genotyping/confirmatory 23S rRNA sequencing can prevent misclassification, reduce unwarranted broad-spectrum escalation, and strengthen antimicrobial stewardship decisions. Full article

Background/Objectives: Small airway dysfunction (SAD) may occur early in children with Mycoplasma pneumoniae pneumonia (MPP), but pulmonary function testing is often deferred until clinical stabilization and may be limited by poor cooperation. Early risk stratification may therefore help identify children who warrant further testing. We aimed to identify early predictors of subsequent spirometry-defined SAD and to develop an internally validated risk-stratification model in hospitalized children with MPP. Methods: In this single-center retrospective cohort study conducted between July 2022 and July 2024, 172 hospitalized children with confirmed MPP were included. Clinical characteristics, immune-inflammatory indices, and chest computed tomography (CT) findings were collected during early hospitalization. Pulmonary function testing was performed after clinical stabilization, and SAD was defined as at least two of forced expiratory flow at 25%, 50%, and 75% of forced vital capacity being <65% of predicted values. Multiple imputation, LASSO selection, and multivariable logistic regression were used for model development and bootstrap internal validation. Results: SAD was identified in 76/172 children (44.2%). Wheezing, CT evidence of small airway involvement, and higher soluble interleukin-2 receptor levels were more common in children with SAD; wheezing remained independently associated with SAD. A model based on routine clinical and chest CT variables showed good discrimination (AUC, 0.885; optimism-corrected AUC, 0.869). Adding interleukin-17 provided limited incremental value. Conclusions: SAD was common in hospitalized children with MPP. An internally validated model based on readily available clinical and chest CT variables may help prioritize children for pulmonary function testing after clinical stabilization, whereas interleukin-17 added limited predictive value. External validation is required before broader clinical application. Full article

Post-infectious bronchiolitis obliterans (PIBO) is a severe chronic airway disease in children following lower respiratory tract infections. Human adenovirus (HAdV) and Mycoplasma pneumoniae (MP) are the major associated pathogens, with geographic variations in their relative importance. This review analytically compares the mechanistic divergence and convergence between HAdV and MP. Both pathogens converge on MyD88/NF-κB/MAPK signaling and neutrophil-driven inflammation, but diverge in initial host engagement (CAR/integrins vs. TLR2/6 and CARDS toxin) and inflammasome activation (TLR9-related vs. NLRP3-related). This review aims to propose an integrative model linking acute immune activation to fibrotic bronchiolar narrowing and to evaluate the risk factors for PIBO. Genetic susceptibility and epigenetic regulation help explain population differences in PIBO risk and geographic distribution. Despite progress, significant knowledge gaps remain, including the lack of single-cell resolution studies, the absence of co-infection animal models, and uncertainty regarding the long-term efficacy of targeted immunomodulatory therapies. Addressing these gaps is essential for improving early diagnosis and clinical outcomes. Full article

Background: Mycoplasma pneumoniae (MP) is an important pathogen responsible for community-acquired respiratory infections in children. Global surveillance during the COVID-19 pandemic revealed a marked decline in MP activity. However, beginning in early summer 2023, multiple regions across China reported an unexpected resurgence of MP infections, highlighting the need for detailed epidemiological analysis. Objective: This study aimed to characterize the epidemiological features of MP seropositivity among children in Chengdu, southwest China, and to compare its patterns between the COVID-19 pandemic and post pandemic periods. Methods: A retrospective analysis was conducted on MP testing data from 39,552 children with acute respiratory infections who were treated at West China Second University Hospital, Sichuan University, between January 2022 and December 2023. Results: Both the number of MP tests conducted and the seropositivity rate were significantly lower during the pandemic period than during the post pandemic phase. Compared with male children, female children were more susceptible to MP seropositivity. In terms of age distribution, seropositivity rates were highest among toddlers (1–3 years) and school-aged children (6–14 years). During the pandemic period (2022), MP antibody-positive cases were observed mainly between January and July, whereas in the post pandemic phase (2023), the epidemic peak shifted from June to December. Conclusions: In this single-centre study in Chengdu, nonpharmaceutical interventions (NPIs) implemented during the COVID-19 pandemic was associated with a marked reduction in MP transmission. After these restrictions were lifted, a rebound in MP antibody positivity was observed among children in Chengdu, compared to the NPI period (2022), the post-NPI period (2023) showed a later seasonal peak, which may represent a delayed return to pre-pandemic patterns. Continuous strengthening of MP surveillance is necessary to provide early warning of potential resurgences and outbreak risks. Full article

Background/Objectives: Mycoplasma pneumoniae (MP) is a frequent cause of community-acquired pneumonia, but it is increasingly recognized for extrapulmonary complications, specifically Mycoplasma pneumoniae-induced rash and mucositis (MIRM). This systematic review aims to comprehensively assess the frequency of clinical features, diagnostic criteria and outcomes of oral mucositis in patients with confirmed MP infection. Methods: A systematic review was conducted following PRISMA guidelines across PubMed, Web of Science and Scopus, covering the period 2015–2025. Inclusion criteria encompassed in vivo studies, case reports, and case series in English focusing on MP-associated mucositis. Methodological quality was assessed using JBI checklists for case-based evidence and the Newcastle–Ottawa Scale for cohort studies. Two clinical cases were reported. Results: Out of 242 identified records, 42 studies were included, involving 140 patients with a notable male predominance (62%). Oral involvement was reported in 92.9% of cases, often characterized by severe ulcerations, hemorrhagic crusting, and debilitating pain. Intensive Care Unit admission was required in 21.5% of cases due to severe systemic or mucosal disease, with 14.3% necessitating parenteral nutrition. Quality assessment indicated moderate-to-high methodological rigor across most included studies. Conclusions: MIRM represents a significant clinical entity where oral mucositis is a dominant feature, often preceding or overshadowing respiratory symptoms. Early recognition by oral health professionals is crucial to avoid misdiagnosis, ensure appropriate multidisciplinary care, and implement supportive or immunomodulatory therapies that reduce morbidity and hospitalization length. Full article

Background/Objectives: This study aimed to investigate the impact of allergic diseases (AD) or obstructive sleep apnea (OSA) risk, as a host factor, on the development of severe Mycoplasma pneumoniae Pneumonia (SMPP) in children by analyzing the clinical data of pediatric patients with Mycoplasma pneumoniae Pneumonia (MPP). Methods: This retrospective study enrolled children hospitalized with Mycoplasma pneumoniae pneumonia (MPP) at Shanghai Ninth People’s Hospital from November 2024 to November 2025. Patients were classified into severe (SMPP) and mild (MMPP) groups. Demographic, clinical, laboratory, and questionnaire data were collected and compared between groups. Univariate and multivariate logistic regression analyses were performed to identify independent predictors of SMPP and construct a nomogram. The model was validated for discrimination, calibration, and clinical utility using ROC curves, calibration plots, and decision curve analysis, with internal validation by bootstrap resampling. Results: Among the 150 enrolled children with MPP, 35 (23.3%) were classified as severe (SMPP) and 115 (76.7%) as mild (MMPP). Patients with SMPP exhibited significantly higher frequencies of allergic diseases, prolonged fever and steroid use, elevated inflammatory markers (CRP, LDH, D-dimer, ferritin, ALT), and higher PSQ and RQLQ scores (all p < 0.05). Disease severity was positively correlated with these clinical, laboratory, and questionnaire-based parameters. Multivariate logistic regression identified allergic diseases, PSQ score, LDH, and ferritin as independent predictors of SMPP. A nomogram incorporating these four factors demonstrated good predictive performance, with an internally validated C-index of 0.827, satisfactory calibration (Hosmer–Lemeshow p = 0.116), and clinical utility within a 0–25% threshold probability range on decision curve analysis. Conclusions: Children with MPP and comorbid AD or OSA risk are more likely to develop SMPP. Among children aged 6–12 years, RQLQ score is positively correlated with the severity of MPP. AD, PSQ score, LDH, and ferritin are independent risk factors for SMPP. Clinicians should be alert to the development of SMPP when children with MPP present with a history of AD, PSQ score >3.5, LDH >327.50 U/L, or ferritin >120.05 ng/mL. The visual nomogram model constructed by combining these risk factors demonstrates improved predictive performance for SMPP, with high predictive efficacy and accuracy. It has great clinical value and can be used for individualized risk assessment and early intervention. However, our proposed nomogram requires external validation prior to broader implementation. Full article

Background: Molecular diagnostics may detect several respiratory pathogens simultaneously with rapid turnaround times. The aim of this study was to determine the frequency and distribution of respiratory pathogens among symptomatic outpatients. Methods: All outpatients presented for testing due to suspected acute respiratory infection between 1 January and 31 December 2024 to the Teaching Institute for Public Health of Split-Dalmatia County, Croatia, and multiplex real-time PCRs for 13 respiratory pathogens were included. Results: Out of 15,437 analyzed panels, 8878 (57.5%) were positive. Single-pathogen infections dominated (82.6%), while co-infections were recorded in 17.4% of panels; therefore, a total of 10,546 individual pathogens were detected, which were mostly viruses (87.0%). The following distribution of pathogens was observed: rhinovirus/enterovirus in 38.9% of positive results, influenza A virus in 14.5%, SARS-CoV-2 in 9.5%, parainfluenza virus in 7.9%, respiratory syncytial virus in 7.3%, Mycoplasma pneumoniae in 4.9%, Bordetella pertussis in 4.6%, human metapneumovirus in 4.2%, adenovirus in 3.4%, Chlamydia pneumoniae in 3.4%, influenza B virus in 1.3%, Bordetella parapertussis in 0.1% and Legionella pneumophila had one positive result. The first trimester of the year had the highest number of positive test panels (47.0%). Conclusions: Our study demonstrates a predominance of viral pathogens across all age groups and seasons, further supporting guideline-based practice and highlighting the importance of confirming bacterial infection before initiating antibiotic therapy. This insight into the post-pandemic circulation of respiratory pathogens may help inform public health strategies, including improved surveillance, anticipation of seasonal outbreaks, and targeted interventions, thereby supporting future pandemic preparedness and mitigation efforts. Full article

Background: Mycoplasma pneumoniae (MP) is a major cause of respiratory tract infections in children and adolescents. Currently, there is no licensed vaccine, underscoring the urgent need for the development of safe and effective vaccines. Objective: The aim of this study is to develop a recombinant subunit vaccine candidate incorporating three antigens: the P1 protein, the P40/90 complex, and a detoxified mutant of community-acquired respiratory distress syndrome toxin. The protective efficacy of this vaccine candidate was also evaluated. Methods: Target genes were codon-optimized for expression in E. coli, and the recombinant proteins were successfully expressed and purified. The low-toxicity CARDS toxin mutant was screened based on TNF-α secretion levels in stimulated RAW264.7 cells. A three-component vaccine composed of P1, P40/90, and the mutant CARDS toxin was formulated and adjuvanted with either Al(OH)₃ alone or in combination with CpG. Mice were immunized, and immunogenicity was assessed by measuring antigen-specific IgG antibody titers. Protective efficacy was evaluated following challenge by analyzing lung histopathology, bacterial load, and inflammatory cytokine levels. Results: Seven high-purity recombinant proteins were successfully produced, including P1, the P40/90 complex, wild-type CARDS toxin, and four CARDS toxin mutants (E132A, E132Q, H36A, R10A). The E132A mutant was selected due to its significantly reduced toxicity while retaining immunogenicity. The three-component vaccine effectively elicited antibody responses against each of the included antigens. After three immunizations, IgG antibody titers in all groups reached approximately 10⁴. Immunized mice showed markedly reduced pulmonary pathology scores (control group: 2 or 2.67; immunized groups: 1.67, 1.33, and 0) and significantly decreased bacterial loads in lung tissue (control: 30.11 ± 10.40 cp/μL; immunized groups: 20.72 ± 4.37 cp/μL and 8.51 ± 8.32 cp/μL). Furthermore, the group receiving the alum + CpG adjuvant exhibited approximately a 10-fold higher antibody response compared with the alum-only group, indicating enhanced protective efficacy. Conclusions: The three-component candidate vaccine, MPtriV, adjuvanted with Al(OH)₃ + CpG, demonstrates promising immunogenicity, safety, and protective efficacy against Mycoplasma pneumoniae infection, providing a viable strategy and experimental foundation for the development of MP subunit vaccines. Full article

Mycoplasma pneumoniae is a significant pathogen responsible for community-acquired respiratory infections in children and adolescents, with the rising prevalence of macrolide-resistant M. pneumoniae (MRMP), particularly in Asia, presenting critical treatment challenges. Our previous study inferred that a macrolide efflux pump may contribute to macrolide resistance in M. pneumoniae in addition to the common point mutations in 23S rRNA gene. This study aimed to define the specific pump and confirm its role. Through comparative genomic analysis, we identified a candidate gene, MPN_080, encoding an ABC transporter permease, which was further characterized using phylogenetic analysis, AlphaFold-based structural modeling, and biochemical assays. Overexpression of MPN_080 from an erythromycin-resistant isolate in the erythromycin-sensitive M129 resulted in a significant increase in minimum inhibitory concentrations (MICs) from <0.125 µg/mL to 1 µg/mL, while similar overexpression of MPN_080 derived from M129 did not affect MICs. Notably, this resistance mechanism operates independently of M. pneumoniae virulence factors, as evidenced by unaltered colonization capacity in NCI-H292 cells and consistent immune response patterns across both strains. Our findings establish MPN_080 as a novel determinant of macrolide resistance functioning associated with enhanced ATPase activity. These insights into non-classical resistance mechanisms may guide future diagnostic and therapeutic strategies against MRMP. Full article

Background: Necrotizing pneumonia is a severe complication of Mycoplasma pneumoniae pneumonia (MPP) in children. Early recognition remains challenging because initial clinical manifestations are often non-specific, highlighting the need for a practical tool for early risk stratification. Methods: We conducted a single-center retrospective study of hospitalized children with MPP. Data from 2015–2023 were used for model development, and patients enrolled in 2024 were reserved for temporal validation. We compared candidate machine-learning algorithms and selected a parsimonious random forest model using routinely available variables obtained during the early hospitalization period. Model performance was evaluated using discrimination, calibration, and decision curve analysis, and model interpretability was assessed using SHapley Additive exPlanations (SHAP). Results: The random forest model showed good discriminatory performance in internal validation and retained acceptable performance in the 2024 temporal cohort. Calibration indicated reasonable agreement between predicted and observed risks. Decision curve analysis suggested potential clinical value as a supportive tool for early risk stratification. SHAP analysis highlighted fever duration, C-reactive protein, pleural effusion, alanine aminotransferase, and gamma-glutamyl transferase as the main contributors to model prediction. Conclusions: We developed and temporally validated a clinical prediction model for necrotizing pneumonia in children hospitalized with MPP. The model may support early risk stratification using routinely available clinical data, but it is intended to complement rather than replace clinical judgment. External prospective validation is required before routine clinical implementation. Full article

Background/Objectives: The aim of this study was to clarify the antimicrobial susceptibility and distribution characteristics of Mycoplasma pneumoniae (M. pneumoniae, MP) collected from children in Beijing, China, from 2017 to 2025. Methods: A total of 197 MP isolates were analyzed. Mutations in macrolide-resistant loci of MP strains were detected via real-time fluorescent quantitative polymerase chain reactions. We used the broth microdilution method to determine the minimum inhibitory concentrations (MICs) of erythromycin, azithromycin, tetracycline, levofloxacin, and moxifloxacin against these isolates. The distribution characteristics of MIC values were further analyzed according to the isolates’ collection year, epidemic phase (low epidemic phase, epidemic initiation phase, ultra-low epidemic phase, outbreak phase, and epidemic recovery phase), and the corresponding patient age group (<3 years, 3–6 years, and ≥6 years). Results: All 197 isolates were found to be resistant to erythromycin and azithromycin, with a resistance rate of 100%. In contrast, the strains remained susceptible to tetracycline, levofloxacin and moxifloxacin. The highest resistance rate was 100% for macrolides. The MIC₉₀ values were 1024 μg/mL for erythromycin, 256 μg/mL for azithromycin, 0.5 μg/mL for tetracycline, 1 μg/mL for levofloxacin, and 0.125 μg/mL for moxifloxacin, respectively. Distinct differences in MIC distributions of erythromycin and azithromycin were observed across collection years, epidemic phases, and age groups. Conclusions: The resistance of MP to macrolides in children is closely associated with the epidemic intensity and age of the patient. Erythromycin is no longer suitable as an empirical therapy for MP infections during epidemic periods, whereas azithromycin can be cautiously administered in young children according to age stratification and MIC detection results. Meanwhile, it is imperative to strengthen the prevention and control of cluster MP infections during epidemic phases to reduce the transmission of drug-resistant MP strains. Full article

Background: To date, no study has compared the clinical characteristics of Mycoplasma pneumoniae-associated segmental/lobar pneumonia, Mycoplasma bronchopneumonia, and COVID-19 pneumonia primarily caused by the NB.1.8.1 variant in children. Methods: We examined the epidemiologic trends of pneumonia, segmental/lobar pneumonia, and COVID-19 pneumonia at a teaching hospital from 2015 to 2025. In addition, we compared the clinical characteristics of children hospitalized with Mycoplasma segmental/lobar pneumonia, Mycoplasma bronchopneumonia, and COVID-19 pneumonia during the NB.1.8.1 variant wave in 2024–2025. Results: Between 2015 and 2024, 10,601 pneumonia cases were identified, including 525 cases of segmental/lobar pneumonia and 162 cases of COVID-19 pneumonia. An outbreak of segmental/lobar M. pneumoniae pneumonia and COVID-19 pneumonia occurred in Taiwan during 2024–2025. Starting in early 2025, monthly Mycoplasma positivity rates among children with segmental/lobar pneumonia and bronchopneumonia exceeded 60%. Mycoplasma pneumonia predominantly affected children aged 6–11 years, whereas COVID-19 pneumonia mainly occurred in those younger than 3 years of age. Fever, cough, and rhinorrhea were the most common symptoms in all groups, limiting clinical differentiation. Children with segmental/lobar Mycoplasma pneumonia were more likely to present with prolonged fever (>5 days), lymphocytopenia, a neutrophil-to-lymphocyte ratio (NLR) ≥ 3, and elevated C-reactive protein (CRP) levels, each of which was strongly associated with macrolide non-response (all p < 0.001). Conclusions: Children with segmental/lobar Mycoplasma pneumonia demonstrated more severe clinical manifestations. Segmental/lobar involvement and inflammatory markers, such as lymphocytopenia, elevated NLR, and increased CRP levels, were associated with macrolide non-response. These indicators may help guide therapeutic decision-making in pediatric M. pneumoniae pneumonia. Full article

The DNA methylation landscape in the lungs of argali hybrid sheep infected with Mycoplasma ovipneumoniae (Mo) remains poorly characterized. This study aimed to profile genome-wide DNA methylation using reduced representation bisulfite sequencing (RRBS) and to validate key genes using bisulfite sequencing PCR (BSP), methylation-specific PCR (MSP), and quantitative MSP (QMSP). The results revealed a significant increase in global mCG methylation in -infected lungs. RRBS identified 3691 differentially methylated regions (DMRs), 66.2% of which were hypermethylated. Methylation levels were highest in gene bodies/downstream regions and lowest in promoters/5′ untranslated regions. Differentially methylated genes (DMGs) were enriched in immune–inflammatory pathways (e.g., antigen presentation, B-cell receptor signaling, Th17 differentiation) and, to a lesser extent, neural signaling pathways. BSP confirmed the methylation status of hypermethylated (KHDC3L, GILT, OVAR-DRB1, SGK1, ADAM17) and hypomethylated (EFCAB11, AP1B1, TATDN1) DMGs. Independent validation by MSP and QMSP further supported the hypermethylation of SGK1 and GILT in both lung tissue and alveolar macrophages. Quantitative reverse-transcription PCR showed that promoter hypermethylation of KHDC3L, GILT, SGK1, and ADAM17 was associated with transcriptional downregulation, while hypomethylation of AP1B1 correlated with upregulation. In summary, Mo infection induces genome-wide hypermethylation reprogramming that dysregulates key immune-related genes, highlighting potential epigenetic mechanisms in the pathogenesis of mycoplasmal pneumonia. Full article

Mycoplasma pneumoniae (M. pneumoniae) is the crucial factor of global acquired respiratory infections. Currently, there are no specific disease modification treatments or vaccines available, and the vaccine development for this pathogen lags behind due to the complexity and variability of its antigens. A novel vaccine with broad-spectrum characteristics is essential to provide comprehensive protection against continuously evolving wild-type strains. Here, a broad-spectrum muti-epitope vaccine against M. pneumoniae had been designed through immunoinformatics methods. To ensure its broad-spectrum, we generated consistent sequences from all the antigen proteins of different strains, and then identified potential T cell epitopes. The multi-epitope vaccine (MEV) of M. pneumoniae incorporated 16 CTLs and 7 HTLs from the HMW1–3 and p1 adhesin proteins, which comprised 458 amino acids with adjuvant. The vaccine evaluation showed that the MEV had ideal physicochemical properties, high antigenicity, high immunogenicity, and was non-toxic. Furthermore, there was a strong and stable binding interaction between this vaccine and the toll-like receptors, which could be supported by the normal mode analysis. Finally, codon optimization resulted in the optimal GC content and higher CAI value. The vaccine candidate is expected to induce strong cellular immune responses and may provide protective immunity against the pathogen. We provided a novel in silico vaccine design strategy for vaccine design, which could provide a technical framework for the development of vaccines against other pathogens. Full article