RESUMO
Mycoplasma pneumoniae can cause respiratory infections and pneumonia, posing a serious threat to the health of children and adolescents. Early diagnosis of Mycoplasma pneumoniae infection is crucial for clinical treatment. Currently, diagnostic methods for Mycoplasma pneumoniae infection include pathogen detection, molecular biology techniques, and bacterial culture, all of which have certain limitations. Here, we developed a rapid, simple, and accurate detection method for Mycoplasma pneumoniae that does not rely on large equipment or complex operations. This technology combines the CRISPR-Cas12a system with recombinase polymerase amplification (RPA), allowing the detection results to be observed through fluorescence curves and immunochromatographic lateral flow strips.It has been validated that RPA-CRISPR/Cas12a fluorescence analysis and RPA-CRISPR/Cas12-immunochromatographic exhibit no cross-reactivity with other common pathogens, and The established detection limit was ascertained to be as low as 102 copies/µL.Additionally, 49 clinical samples were tested and compared with fluorescence quantitative polymerase chain reaction, demonstrating a sensitivity and specificity of 100%. This platform exhibits promising clinical performance and holds significant potential for clinical application, particularly in settings with limited resources, such as clinical care points or resource-constrained areas.
Assuntos
Sistemas CRISPR-Cas , Mycoplasma pneumoniae , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/isolamento & purificação , Humanos , Sistemas CRISPR-Cas/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/microbiologiaRESUMO
Introduction. Recently, the incidence of Mycoplasma pneumoniae (M. pneumoniae) infection in children has been increasing annually. Early differential diagnosis of M. pneumoniae infection can not only avoid the abuse of antibiotics, but also is essential for early treatment and reduction of transmission.Gap statement. The change of routine blood parameters may have important clinical significance for the diagnosis of M. pneumoniae infection, but it has not been reported so far.Aim. This study aims to establish a predictive model for M. pneumoniae infection and explore the changes and clinical value of routine blood parameters in children with M. pneumoniae infection, serving as auxiliary indicators for the diagnosis and differentiation of clinical M. pneumoniae infection.Methodology. A total of 770 paediatric patients with respiratory tract infections were enrolled in this study, including 360 in the M. pneumoniae group, 40 in the SARS-CoV-2 group, 200 in the influenza A virus group, and 170 in the control group. The differences of routine blood parameters among all groups were compared, and risk factors were analysed using multivariate logistics analysis, and the diagnostic efficacy of differential indicators using ROC curves.Results. This study revealed that Mono% (OR: 3.411; 95% CI: 1.638-7.102; P=0.001) was independent risk factor associated with M. pneumoniae infection, and Mono% (AUC=0.786, the optimal cutoff at 7.8%) had a good discriminative ability between patients with M. pneumoniae infection and healthy individuals. Additionally, Mono% (OR: 0.424; 95% CI: 0.231-0.781; P=0.006) and Lymp% (OR: 0.430; 95% CI: 0.246-0.753; P=0.003) were independent risk factors for distinguishing M. pneumoniae infection from influenza A virus infection, and the Lymp% (AUC=0.786, the optimal cutoff at 22.1%) and Net% (AUC=0.761, the optimal cutoff at 65.2%) had good discriminative abilities between M. pneumoniae infection and influenza A infection. Furthermore, platelet distribution width (OR: 0.680; 95% CI: 0.538-0.858; P=0.001) was independent risk factor for distinguishing M. pneumoniae infection from SARS-CoV-2 infection. Meanwhile, the ROC curve demonstrated that PDW (AUC=0.786, the optimal cutoff at 15%) has a good ability to differentiate between M. pneumoniae infection and SARS-CoV-2 infection.Conclusion. This study demonstrates that routine blood parameters can be used as auxiliary diagnostic indicators for M. pneumoniae infection and provide reference for the diagnosis and differentiation of clinical M. pneumoniae infection.
Assuntos
Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/microbiologia , Feminino , Masculino , Pré-Escolar , Criança , Mycoplasma pneumoniae/isolamento & purificação , COVID-19/diagnóstico , COVID-19/sangue , Lactente , Curva ROC , Fatores de Risco , Diagnóstico Diferencial , Adolescente , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/microbiologia , Infecções Respiratórias/sangue , SARS-CoV-2/isolamento & purificaçãoRESUMO
BACKGROUND: Blood routine testing was the most commonly used laboratory method in clinical practice. The results are often influenced by factors such as instruments, reagents, and samples, among which, the interference of cold agglutinin is a very rare element. In our article, we reported a case of red blood cell agglutination caused by Mycoplasma pneumoniae infection. METHODS: The number of blood cells were detected by blood routine analyzer with or without treatment at 37â water bath. The red blood cell agglutination was observed through blood smear staining. The cold agglutination test were performed using O-type red blood cells added into patient's plasma and refrigerated overnight at 4â. We also used luminescent immunoassay technology to detect the content of MP antibodies in patient's serum. RESULTS: The patient's results were RBC (2.69 x 1012/L), MCH (48.5 pg), MCHC (522 g/L). Through a microscope, we observed red blood cell agglutination. The concentration of MP-igM was 60.37 AU/mL. The cold agglutination test was positive. Following a 37â water bath, the patient's results changed: RBC (3.85 x 1012/L), MCH (31.2 pg), MCHC (352 g/L). The phenomenon of massive agglutination of red blood cells has also disappeared. CONCLUSIONS: The cold agglutinin produced by MP infection can alter the results of red blood cell. During the epidemic period of MP infection, it is important to pay attention to the phenomenon of abnormal elevation of MCHC in clinical practice.
Assuntos
Eritrócitos , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/microbiologia , Mycoplasma pneumoniae/imunologia , Crioglobulinas/análise , Crioglobulinas/metabolismo , Masculino , Testes de Aglutinação , Aglutinação , Feminino , Imunoglobulina M/sangueRESUMO
BACKGROUND: Mycoplasma pneumoniae (MP) is a common respiratory pathogen affecting the longevity of the elderly and the health of children. However, the human vaccine against MP has not been successfully developed till now due to the poor immunogenicity and side effects of MP inactivated or attenuated vaccine. Therefore, it is necessary to develop a MP genetic engineering vaccine with influenza virus strain as vector. METHODS: In this study, the major antigen genes P1a of MP adhesion factor P1(3862-4554 bases) and P30a of P30(49-822 bases) were inserted into the nonstructural protein (NS) gene of Influenza A virus strain A/Puerto Rio/8/34(H1N1), PR8 for short, to construct the recombinant vectors NS-P1a or NS-P30a. The recombinant pHW2000 plasmids containing NS-P1a or NS-P30a were cotransfected with the rest 7 fragments of PR8 into HEK293T cells. After inoculating chicken embryos, the recombinant influenza viruses rFLU-P1a and rFLU-P30a were rescued. RT-PCR and sequencing were used to identify the recombinant viruses. The hemagglutination titers of rFLU-P1a and rFLU-P30a were determined after five successive generations in chicken embryos so as to indicate the genetic stability of the recombinant viruses. The morphology of recombinant influenza viruses was observed under electron microscopy. RESULTS: P1a or P30a was designed to be inserted into the modified NS gene sequence separately and synthesized successfully. RT-PCR identification of the recombinant viruses rFLU-P1a and rFLU-P30a showed that P1a (693 bp), P30a (774 bp), NS-P1a (1992bp) and NS-P30a (2073 bp) bands were found, and the sequencing results were correct. After five successive generations, each virus generation has a certain hemagglutination titer (from 1:32 to 1:64), and the band of P1a or P30a can be seen in the corresponding positions. The virus particles under the electron microscope appeared as spheres or long strips connected by several particles, revealing a complete viral membrane structure composed of virus lipid bilayer, hemagglutinin, neuraminidase, and matrix proteins. CONCLUSION: The recombinant viruses rFLU-P1a and rFLU-P30a which carried the advantaged immune regions of the P1 and P30 genes in MP were successfully constructed and identified. And the genetic stability of rFLU-P1a or rFLU-P30a was relatively high. The typical and complete morphology of influenza virus was observed under the electron microscope. Our research provided a foundation for the further development of MP vaccines for human.
Assuntos
Vetores Genéticos , Mycoplasma pneumoniae , Humanos , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/imunologia , Animais , Células HEK293 , Vetores Genéticos/genética , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/imunologia , Adesinas Bacterianas/genética , Adesinas Bacterianas/imunologia , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/imunologia , Vírus da Influenza A/genética , Vírus da Influenza A/imunologia , Embrião de Galinha , Pneumonia por Mycoplasma/imunologia , Vacinas Sintéticas/imunologia , Vacinas Sintéticas/genéticaRESUMO
BACKGROUND: This study aims to report the phenomenon of Myelin oligodendrocyte glycoprotein antibody-associated encephalitis induced by Mycoplasma pneumoniae infections and promote the potential benefits of combining early immunotherapy and anti-M-pneumoniae therapy for these patients. METHODS: Three children with MOG-IgG-associated encephalitis due to M. pneumoniae infections who were treated at our hospital from September to November 2023 were included in the study. We investigated and analyzed the background and clinical features of these patients. RESULTS: Three patients developed headaches, seizures, and/or other neurological manifestations, elevated mononuclear cells in cerebrospinal fluid, intracranial lesions on cranial magnetic resonance imaging (MRI), and positive MOG-IgG in serum, within 10-14 days. They were diagnosed with MOG-IgG-associated encephalitis due to M. pneumoniae infections, the treatment consisted of intravenous immunoglobulin, glucocorticoid, and erythromycin, then they were completely recovered. CONCLUSION: Mycoplasma pneumoniae (M. pneumoniae) infections can cause oligodendrocyte glycoprotein (MOG) antibody-associated encephalitis. The recognition of this condition will promote the potential benefits of combining early immunotherapy and anti-M. pneumoniae therapy for patients with MOG-IgG-associated encephalitis.
Assuntos
Mycoplasma pneumoniae , Glicoproteína Mielina-Oligodendrócito , Pneumonia por Mycoplasma , Humanos , Glicoproteína Mielina-Oligodendrócito/imunologia , Pneumonia por Mycoplasma/complicações , Pneumonia por Mycoplasma/diagnóstico , Masculino , Feminino , Criança , Mycoplasma pneumoniae/imunologia , Pré-Escolar , Encefalite/imunologia , Encefalite/diagnóstico , Imunoglobulina G/sangue , Autoanticorpos/sangue , Imageamento por Ressonância MagnéticaRESUMO
BACKGROUND: X-linked agammaglobulinemia (XLA), also referred to as Bruton's tyrosine kinase deficiency, is a rare genetic disorder that affects the immune system. We conducted genetic analysis on patients suffering from immunodeficiency by utilizing Next-Generation Sequencing techniques, as well as their closest relatives, to facilitate accurate diagnosis, offer genetic counseling services, and enhance our comprehension of XLA.
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Agamaglobulinemia , Doenças Genéticas Ligadas ao Cromossomo X , Pneumonia por Mycoplasma , Humanos , Agamaglobulinemia/complicações , Agamaglobulinemia/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Doenças Genéticas Ligadas ao Cromossomo X/complicações , Masculino , Pneumonia por Mycoplasma/complicações , Pneumonia por Mycoplasma/microbiologia , Tirosina Quinase da Agamaglobulinemia/genética , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/isolamento & purificação , Adulto , Sequenciamento de Nucleotídeos em Larga EscalaRESUMO
BACKGROUND: Mycoplasmal pneumonia of sheep and goats (MPSG) is an important infectious disease that threatens sheep and goat production worldwide, and Mycoplasma ovipneumoniae (Movi) is one of the major aetiological agents causing MPSG. The aim of this study was to investigate the immunological activity of the Hsp70âP113 fusion protein derived from Movi and to develop a serological assay for the detection of Movi. METHODS: This study involved codon optimization of the dominant antigenic regions of Movi heat shock protein 70 (Hsp70) and adhesin P113. Afterwards, the optimized sequences were inserted into the prokaryotic expression vector pET-30a( +) through tandem linking with the aid of a linker. Once a positive recombinant plasmid (pET-30a-rHsp70-P113) was successfully generated, the expression conditions were further refined. The resulting double gene fusion target protein (rHsp70âP113) was subsequently purified using ProteinIso® Ni-NTA resin, and the reactivity of the protein was confirmed via SDSâPAGE and Western blot analysis. An indirect enzyme-linked immunosorbent assay (i-ELISA) technique was developed to detect Movi utilizing the fusion protein as the coating antigen. The specificity, sensitivity, and reproducibility of all methods were assessed after each reaction parameter was optimized. RESULTS: The resulting rHsp70-P113 protein had a molecular weight of approximately 51 kDa and was predominantly expressed in the supernatant. Western blot analysis demonstrated its favourable reactivity. The optimal parameters for the i-ELISA technique were as follows: the rHsp70-P113 protein was encapsulated at a concentration of 5 µg/mL; the serum was diluted at a ratio of 1:50; the HRP-labelled donkey anti-goat IgG was diluted at a ratio of 1:6,000. The results of the cross-reactivity assays revealed that the i-ELISA was not cross-reactive with other goat-positive sera against Mycoplasma mycodies subsp. capri (Mmc), Mycoplasma capricolum subsp. capripneumoniae (Mccp), Mycoplasma arginini (Marg), orf virus (ORFV) or enzootic nasal tumour virus of goats (ENTV-2). The sensitivity of this method is high, with a maximum dilution of up to 1:640. The results of the intra- and inter-batch replication tests revealed that the coefficients of variation were both less than 10%, indicating excellent reproducibility. The analysis of 108 clinical serum samples via i-ELISA and indirect haemagglutination techniques yielded significant findings. Among these samples, 43 displayed positive results, whereas 65 presented negative results, resulting in a positivity rate of 39.8% for the i-ELISA method. In contrast, the indirect haemagglutination technique identified 20 positive samples and 88 negative samples, resulting in a positivity rate of 18.5%. Moreover, a comparison between the two methods revealed a conformity rate of 78.7%. CONCLUSION: The results obtained in this study lay the groundwork for advancements in the use of an Movi antibody detection kit, epidemiological inquiry, and subunit vaccines.
Assuntos
Ensaio de Imunoadsorção Enzimática , Doenças das Cabras , Cabras , Proteínas de Choque Térmico HSP70 , Mycoplasma ovipneumoniae , Pneumonia por Mycoplasma , Proteínas Recombinantes de Fusão , Doenças dos Ovinos , Animais , Mycoplasma ovipneumoniae/imunologia , Mycoplasma ovipneumoniae/genética , Proteínas de Choque Térmico HSP70/imunologia , Proteínas de Choque Térmico HSP70/genética , Doenças das Cabras/diagnóstico , Doenças das Cabras/imunologia , Doenças das Cabras/microbiologia , Ensaio de Imunoadsorção Enzimática/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/microbiologia , Ovinos , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Pneumonia por Mycoplasma/veterinária , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/imunologia , Adesinas Bacterianas/imunologia , Adesinas Bacterianas/genética , Anticorpos Antibacterianos/sangue , Sensibilidade e Especificidade , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/genéticaRESUMO
OBJECTIVES: To explore the establishment of a risk prediction model for concurrent bronchiolitis obliterans (BO) in children with refractory Mycoplasma pneumoniae pneumonia (RMPP). METHODS: A retrospective study included 116 RMPP children treated in the Department of Pediatrics of Xiangya Changde Hospital from June 2021 to December 2023. Eighty-one cases were allocated to the training set and thirty-five cases to the validation set based on a 7:3 ratio. Among them, 26 cases in the training set developed BO, while 55 did not. The multivariate logistic regression was used to select variable factors for constructing the BO risk prediction model. Nomograms were drawn, and the receiver operating characteristic (ROC) curve was used to assess the discriminative ability of the model, while calibration curves and Hosmer-Lemeshow tests evaluated the model's calibration. RESULTS: Multivariate logistic regression analysis indicated that several factors were significantly associated with concurrent BO in RMPP children, including length of hospital stay, duration of fever, atelectasis, neutrophil percentage (NEUT%), peak lactate dehydrogenase (LDH), ferritin, peak C reactive protein (CRP), oxygenation index (PaO2/FiO2), ≥2/3 lung lobe consolidation, pleural effusion, bronchial mucous plugs, bronchial mucosal necrosis, and arterial oxygen partial pressure (PaO2) (P<0.05). ROC curve analysis for the training set indicated an area under the curve of 0.904 with 88% sensitivity and 83% specificity; the validation set showed an area under the curve of 0.823 with 76% sensitivity and 93% specificity. The Hosmer-Lemeshow test's Chi-square values for the training and validation sets were 2.17 and 1.92, respectively, with P values of 0.221 and 0.196, respectively. CONCLUSIONS: The risk prediction model for BO in RMPP children based on logistic regression has good performance. Variables such as length of hospital stay, duration of fever, atelectasis, peak LDH, peak CRP, NEUT%, ferritin, ≥2/3 lung lobe consolidation, pleural effusion, bronchial mucous plugs, bronchial mucosal necrosis, PaO2/FiO2, andPaO2 can be used as predictors.
Assuntos
Bronquiolite Obliterante , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/complicações , Feminino , Masculino , Estudos Retrospectivos , Criança , Modelos Logísticos , Bronquiolite Obliterante/etiologia , Pré-Escolar , Curva ROC , NomogramasRESUMO
Background: Mycoplasma pneumoniae (MP) is a significant cause of community-acquired pneumonia with high macrolide resistance rates. Various COVID-19 pandemic restrictions have impacted the prevalence of MP. Objective: To assess the changes in the pattern of MP infections among children before, during, and after the COVID-19 pandemic. Methods: A total of 36685 enrolled patients, aged 0-18 years, diagnosed with pneumonia and admitted to Children's Hospital of Chongqing Medical University from January 2019 to December 2023, were retrospectively reviewed in this study. The epidemiological characteristics of pediatric MP infection were analyzed. Results: Among 36685 patients, 7610 (20.74%) tested positive for MP. The highest positive rate was observed among children aged over 6 years (55.06%). There was no gender disparity in MP infection across the three phases of the COVID-19 pandemic. Hospital stays were longest for children during the COVID-19 pandemic (P <0.001). MP infection was most prevalent in the summer (29.64%). The lowest positive rate was observed during the pandemic, with the highest rate found after easing the measures across all age groups (P <0.001). There was a surge in the positive rate of MP in the third year after the COVID-19 pandemic. Regression analyses demonstrated a shift in the age range susceptible to MP infection, with children aged 3.8 to 13.5 years post-pandemic compared to the pre-pandemic range of 5.3 to 15.5 years old. Additionally, the average macrolide resistance rate was 79.84%. We observed a higher resistance rate during the pandemic than in the pre- and post-pandemic phases (P <0.001). Conclusion: The restrictive measures implemented during the COVID-19 pandemic have influenced the spread of MP to some extent and altered demographic and clinical characteristics, such as age, age group, season, length of stay, and macrolide resistance. We recommend continuous surveillance of the evolving epidemiological characteristics of MP infection in the post-pandemic period when restrictions are no longer necessary.
Assuntos
COVID-19 , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , SARS-CoV-2 , Humanos , Criança , COVID-19/epidemiologia , Pré-Escolar , China/epidemiologia , Feminino , Masculino , Lactente , Adolescente , Pneumonia por Mycoplasma/epidemiologia , Mycoplasma pneumoniae/efeitos dos fármacos , Mycoplasma pneumoniae/isolamento & purificação , Estudos Retrospectivos , Recém-Nascido , Prevalência , SARS-CoV-2/isolamento & purificação , Hospitalização/estatística & dados numéricos , Pandemias , Macrolídeos/uso terapêutico , Farmacorresistência Bacteriana , Antibacterianos/uso terapêutico , Antibacterianos/farmacologia , Criança Hospitalizada/estatística & dados numéricos , Estações do Ano , Infecções Comunitárias Adquiridas/epidemiologia , Infecções Comunitárias Adquiridas/microbiologiaRESUMO
BACKGROUND: The clinical significance of the presence or absence of Mycoplasma pneumoniae (MP) in pleural effusion in Mycoplasma pneumoniae pneumonia (MPP) children has not yet been elucidated. Herein, we investigated the clinical implication of pleural fluid MP positive in children with MPP. METHODS: A total of 165 MPP children with pleural effusion requiring thoracocentesis were enrolled in this study. They were subsequently divided into two groups according to the presence or absence of MP in pleural effusion, namely positive group (n = 38) and negative group (n = 127). Information on their clinical manifestations, laboratory findings, radiological characteristics and treatment modalities was retrospectively collected from medical chart reviews. RESULTS: The length of hospitalization (15.00 (10.75-19.25) vs. 11.00 (9.00-14.00) days, p=0.001) and total course of illness (23.00 (18.00-28.00) vs. 20.00 (17.00-24.00) days, p=0.010) were significantly longer in the positive group than in the negative group. The occurrence of pericardial effusion (23.7% vs. 7.9%, p=0.017), atelectasis (73.7% vs. 53.5%, p=0.027) and necrotizing pneumonia (23.7% vs. 7.9%, p=0.017) were more frequent in the positive group compared to the negative group. The levels of neutrophil percentages (82.35% (75.40%-85.78%) vs. 72.70% (64.30%-79.90%), p<0.001), C-reactive protein (CRP) (71.12 (37.75-139.41) vs. 31.15 (13.54-65.00) mg/L, p<0.001), procalcitonin (PCT) (0.65 (0.30-3.05) vs. 0.33 (0.17-1.13) ng/ml, p=0.005), serum lactate dehydrogenase (LDH) (799.00 (589.00-1081.50) vs. 673.00 (503.00-869.00) U/L, p=0.009), D-dimer (6.21 (3.37-16.11) vs. 3.32 (2.12-6.62) mg/L, p=0.001) on admission were significantly higher in the positive group than in the negative group. These pronounced differences significantly contributed to the identification of MPP with MP positive pleural effusion, as evidenced by the ROC curve analysis. Marked elevations in adenosine deaminase (49.25 (36.20-60.18) vs. 36.20 (28.10-46.50) U/L, p<0.001) and LDH levels (2298.50 (1259.75-3287.00) vs. 1199.00 (707.00-1761.00) U/L, p<0.001) were observed in pleural fluid of the positive group when compared to the negative group. Meanwhile, the number of patients on low molecular weight heparin (LMWH) therapy (9 (23.7%) vs. 12 (9.4%), p=0.028) was higher in the positive group. Multivariate logistic regression analysis revealed that D-dimer > 7.33 mg/L was significantly associated with the incidence of MP positive pleural effusion in MPP (OR=3.517). CONCLUSIONS: The presence of MP in pleural fluid in MPP children with pleural effusion indicated a more serious clinical course. D-dimer > 7.33 mg/L was a related factor for MP positive pleural effusion in MPP. The results of the present study would help in the creation of a therapeutic plan and prediction of the clinical course of MPP in children.
Assuntos
Mycoplasma pneumoniae , Derrame Pleural , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/epidemiologia , Feminino , Estudos Retrospectivos , Derrame Pleural/microbiologia , Masculino , Pré-Escolar , Criança , Lactente , Proteína C-Reativa/análise , Tempo de InternaçãoRESUMO
BACKGROUND: Mycoplasma pneumoniae pneumonia (MPP) is responsible for 20 to 40% of all cases of pneumonia acquired by children and shows an increasing incidence year by year. The aim of this study was to investigate the expression of miR-34a in children with MPP and its diagnostic value, and further explore the relationship between miR-34a and the rehabilitation effect of children with MPP. METHODS: The expression level of miR-34a was detected by RT-qPCR, and the clinical value of miR-34a was analyzed by ROC analysis. In addition, the levels of IL-6, IL-18 and TNF-α in serum of children with MPP were detected by ELISA kit, and the correlation with miR-34a was analyzed. RESULTS: Elevated levels of miR-34a were observed in the serum of children with MPP, and significantly higher expression levels were observed in children with severe symptoms and poor rehabilitation. The study suggested that miR-34a has potential as a diagnostic marker for MPP in children, helping to distinguish between mild and severe cases and predicting rehabilitation from MPP in children. In addition, miR-34a expression was positively correlated with IL-6, IL-8, and TNF-α levels. CONCLUSIONS: miR-34a is closely related to MPP in children and miR-34a may be used as a clinical biomarker for MPP in children.
Assuntos
MicroRNAs , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/sangue , Masculino , Feminino , MicroRNAs/sangue , Criança , Pré-Escolar , Biomarcadores/sangue , Mycoplasma pneumoniae/genéticaRESUMO
The COVID-19 pandemic has underscored the critical need for precise diagnostic methods to distinguish between similar respiratory infections, such as COVID-19 and Mycoplasma pneumoniae (MP). Identifying key biomarkers and utilizing machine learning techniques, such as random forest analysis, can significantly improve diagnostic accuracy. We conducted a retrospective analysis of clinical and laboratory data from 214 patients with acute respiratory infections, collected between October 2022 and October 2023 at the Second Hospital of Nanping. The study population was categorized into three groups: COVID-19 positive (n = 52), MP positive (n = 140), and co-infected (n = 22). Key biomarkers, including C-reactive protein (CRP), procalcitonin (PCT), interleukin- 6 (IL-6), and white blood cell (WBC) counts, were evaluated. Correlation analyses were conducted to assess relationships between biomarkers within each group. The random forest analysis was applied to evaluate the discriminative power of these biomarkers. The random forest model demonstrated high classification performance, with area under the ROC curve (AUC) scores of 0.86 (95% CI: 0.70-0.97) for COVID-19, 0.79 (95% CI: 0.64-0.92) for MP, 0.69 (95% CI: 0.50-0.87) for co-infections, and 0.90 (95% CI: 0.83-0.95) for the micro-average ROC. Additionally, the precision-recall curve for the random forest classifier showed a micro-average AUC of 0.80 (95% CI: 0.69-0.91). Confusion matrices highlighted the model's accuracy (0.77) and biomarker relationships. The SHAP feature importance analysis indicated that age (0.27), CRP (0.25), IL6 (0.14), and PCT (0.14) were the most significant predictors. The integration of computational methods, particularly random forest analysis, in evaluating clinical and biomarker data presents a promising approach for enhancing diagnostic processes for infectious diseases. Our findings support the use of specific biomarkers in differentiating between COVID-19 and MP, potentially leading to more targeted and effective diagnostic strategies. This study underscores the potential of machine learning techniques in improving disease classification in the era of precision medicine.
Assuntos
Biomarcadores , Proteína C-Reativa , COVID-19 , Aprendizado de Máquina , Pneumonia por Mycoplasma , Pró-Calcitonina , Humanos , COVID-19/diagnóstico , COVID-19/sangue , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/sangue , Biomarcadores/sangue , Masculino , Feminino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto , Diagnóstico Diferencial , Pró-Calcitonina/sangue , Proteína C-Reativa/análise , Proteína C-Reativa/metabolismo , Idoso , Interleucina-6/sangue , SARS-CoV-2/isolamento & purificação , Coinfecção/diagnóstico , Coinfecção/sangue , Mycoplasma pneumoniae , Contagem de Leucócitos , Curva ROC , Algoritmo Florestas AleatóriasRESUMO
OBJECTIVES: Mycoplasma pneumoniae (M. pneumoniae) continues to pose a significant disease burden on global public health as a respiratory pathogen. The antimicrobial resistance among M. pneumoniae strains has complicated the outbreak control efforts, emphasizing the need for robust surveillance systems and effective antimicrobial stewardship programs. DESIGN: This review comprehensively investigates studies stemming from previous outbreaks to emphasize the multifaceted nature of M. pneumoniae infections, encompassing epidemiological dynamics, diagnostic innovations, antibiotic resistance, and therapeutic challenges. RESULTS: We explored the spectrum of clinical manifestations associated with M. pneumoniae infections, emphasizing the continuum of disease severity and the challenges in gradating it accurately. Artificial intelligence and machine learning have emerged as promising tools in M. pneumoniae diagnostics, offering enhanced accuracy and efficiency in identifying infections. However, their integration into clinical practice presents hurdles that need to be addressed. Further, we elucidate the pivotal role of pharmacological interventions in controlling and treating M. pneumoniae infections as the efficacy of existing therapies is jeopardized by evolving resistance mechanisms. CONCLUSION: Lessons learned from previous outbreaks underscore the importance of adaptive treatment strategies and proactive management approaches. Addressing these complexities demands a holistic approach integrating advanced technologies, genomic surveillance, and adaptive clinical strategies to effectively combat this pathogen.
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Antibacterianos , Surtos de Doenças , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Mycoplasma pneumoniae/efeitos dos fármacos , Mycoplasma pneumoniae/genética , Pneumonia por Mycoplasma/epidemiologia , Pneumonia por Mycoplasma/tratamento farmacológico , Pneumonia por Mycoplasma/microbiologia , Pneumonia por Mycoplasma/diagnóstico , Antibacterianos/uso terapêutico , Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Inteligência Artificial , Gestão de AntimicrobianosRESUMO
Introduction: Mycoplasma pneumoniae (MP) is the major cause of respiratory infections that threaten the health of children and adolescents worldwide. Therefore, an early, simple, and accurate detection approach for MP is critical to prevent outbreaks of MP-induced community-acquired pneumonia. Methods: Here, we explored a simple and accurate method for MP identification that combines loop-mediated isothermal amplification (LAMP) with the CRISPR/Cas12b assay in a one-pot reaction. Results: In the current study, the whole reaction was completed within 1 h at a constant temperature of 57°C. The limit of detection of this assay was 33.7 copies per reaction. The specificity of the LAMP-CRISPR/Cas12b method was 100%, without any cross-reactivity with other pathogens. Overall, 272 clinical samples were used to evaluate the clinical performance of LAMP-CRISPR/Cas12b. Compared with the gold standard results from real-time PCR, the present method provided a sensitivity of 88.11% (126/143), specificity of 100% (129/129), and consistency of 93.75% (255/272). Discussion: Taken together, our preliminary results illustrate that the LAMP-CRISPR/Cas12b method is a simple and reliable tool for MP diagnosis that can be performed in resource-limited regions.
Assuntos
Sistemas CRISPR-Cas , Técnicas de Diagnóstico Molecular , Mycoplasma pneumoniae , Técnicas de Amplificação de Ácido Nucleico , Pneumonia por Mycoplasma , Sensibilidade e Especificidade , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Humanos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/microbiologia , Técnicas de Diagnóstico Molecular/métodos , Criança , Limite de DetecçãoRESUMO
BACKGROUND: This study investigated clinical and laboratory characteristics of human bocavirus type 1 (HBoV1)-plastic bronchiolitis (PB), Mycoplasma pneumoniae (MP)-associated plastic bronchitis (PB) and MP-NPB in children, highlighting inflammation, coagulation, and bronchoscopic needs. METHODS: Data on preschool children with PB during HBoV1 or MP infection were collected, comparing MP-PB to severe Mycoplasma pneumoniae pneumonia. RESULT: Compared with the MP-PB group, the HBoV1-PB group, with younger children, had significantly milder clinical symptoms but higher WBC counts (p = .028). The MP-PB group exhibited notably elevated Fibrinogen (p = .045) and d-dimer levels (p < .001). When contrasting the MP-PB with the MP-NPB group, children in MP-PB group still had higher levels of d-dimer and increased inflammatory indicators such as C-reactive protein, procalcitonin, lactate dehydrogenase, and interleukin-6, which were significantly elevated compared with the MP-NPB group. MP-PB showed a higher prevalence of plastic bronchial casts in lower lobes (p = .016) and a dominance of neutrophils in BALF cytology. Additionally, children in the MP-PB group tended to undergo a greater number of bronchoscopies. CONCLUSION: This study identifies key differences in plastic bronchitis in children due to HBoV1 and MP, highlighting HBoV1's milder inflammation in younger kids and MP's link to severe inflammatory and coagulation responses, guiding clinical diagnosis and treatment.
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Bronquite , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pré-Escolar , Masculino , Feminino , Bronquite/microbiologia , Bronquite/diagnóstico , Bronquite/virologia , Pneumonia por Mycoplasma/sangue , Pneumonia por Mycoplasma/imunologia , Lactente , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/complicações , Infecções por Parvoviridae/diagnóstico , Bocavirus Humano , Bronquiolite/virologia , Bronquiolite/microbiologia , Criança , Líquido da Lavagem Broncoalveolar/virologia , Líquido da Lavagem Broncoalveolar/microbiologia , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , Proteína C-Reativa/análiseRESUMO
Since October 2023, a significant outbreak of Mycoplasma Pneumoniae Pneumonia (MPP) has been observed in children in northern China. Chinese health authorities have attributed this epidemiological to immune debt resulting from the relaxation of coronavirus disease 2019 (COVID-19) control measures. This study described the epidemiological features of Mycoplasma pneumoniae (MP) prevalence in children and developed a straightforward prediction model to differentiate between MPP and viral pneumonia in children. The infection rate of MP in children notably increased from 8.12 in 2022 to 14.94% in 2023, peaking between October and November, especially among school-age children. Logistic regression screening identified four key indicators: Age, D-Dimer levels, erythrocyte sedimentation rate, and gender. The developed nomogram exhibited a receiver operator characteristic curve-area under the curve (ROC-AUC) of 0.858, with external validation confirming an ROC-AUC of 0.794. This study examined the epidemiological characteristics of MPP prevalence in children in Shandong Province during and after the COVID-19 pandemic. An early predict model was developed and validated to differentiate between Mycoplasma Pneumoniae and viral infections.
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COVID-19 , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/epidemiologia , COVID-19/epidemiologia , China/epidemiologia , Criança , Feminino , Masculino , Pré-Escolar , Adolescente , Lactente , Surtos de Doenças , Prevalência , SARS-CoV-2/isolamento & purificação , Curva ROCRESUMO
Community-Acquired Respiratory Distress Syndrome Toxin (CARDS TX) is a unique exotoxin produced by Mycoplasma pneumoniae (MP) and has been confirmed to possess ADP-ribosyltransferase (ART) and vacuolating activities. CARDS TX binds to receptors on the surfaces of mammalian cells followed by entry into the cells through clathrin-mediated endocytosis, and exerts cytotoxic effects by undergoing retrograde transport and finally cleavage on endosomes and cellular organelles. In addition, CARDS TX can trigger severe inflammatory reactions resulting in airway dysfunction, producing allergic inflammation and asthma-like conditions. As a newly discovered virulence factor of MP, CARDS TX has been extensively studied in recent years. As resistance to macrolide drugs has increased significantly in recent years and there is no vaccine against MP, the development of a vaccine targeting CARDS TX is considered a potential preventive measure. This review focuses on recent studies and insights into this toxin, providing directions for a better understanding of MP pathogenesis and treatment. IMPORTANCE: A serious hazard to worldwide public health in recent years, Mycoplasma pneumoniae (MP) is a prominent bacterium that causes community-acquired pneumonia (CAP) in hospitalized children. Due to their high prevalence and fatality rates, MP infections often cause both respiratory illnesses and extensive extrapulmonary symptoms. It has recently been shown that MP produces a distinct exotoxin known as Community-Acquired Respiratory Distress Syndrome Toxin (CARDS TX). Mycoplasma pneumoniae pneumonia (MPP)-like tissue injury is caused by this toxin because it has both ADP-ribosyltransferase and vacuolating properties. A better knowledge of MP etiology and therapy is provided by this review, which focuses on latest research and insights into this toxin.
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ADP Ribose Transferases , Proteínas de Bactérias , Toxinas Bacterianas , Infecções Comunitárias Adquiridas , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Fatores de Virulência , Mycoplasma pneumoniae/patogenicidade , Humanos , Infecções Comunitárias Adquiridas/microbiologia , Toxinas Bacterianas/metabolismo , Pneumonia por Mycoplasma/microbiologia , Fatores de Virulência/metabolismo , ADP Ribose Transferases/metabolismo , Proteínas de Bactérias/metabolismo , Animais , Virulência , Exotoxinas/metabolismo , Síndrome do Desconforto Respiratório/microbiologia , Síndrome do Desconforto Respiratório/etiologiaRESUMO
BACKGROUND: This study aims to analyse changes in urinary kidney injury markers in children with Mycoplasma pneumoniae pneumonia (MPP), investigate the risk factors for MPP-related acute kidney injury (AKI) and establish a model to predict MPP-related AKI. METHODS: Ninety-five children were enrolled based on the study's inclusion and exclusion criteria. They were divided into a severe MPP (SMPP) group and a non-SMPP group and then into an AKI group and a non-AKI group according to the presence of AKI. A univariate logistic regression analysis was performed to explore the early risk factors for AKI. Based on a multivariate logistic regression analysis and a least absolute shrinkage and selection operator regression analysis, appropriate variables were selected to establish a prediction model, and R 4.2.2 software was used to draw nomograms and generate a dynamic nomogram website. RESULTS: Seven urinary kidney injury markers were abnormally elevated in the SMPP group and the non-SMPP group: urinary N-acetyl-ß-D-glucosaminidase (NAG), ß2-microglobulin, α1-microglobulin, retinol-binding protein, urinary immunoglobulin G, urinary transferrin and urinary microalbumin. Sixteen children were identified with AKI during hospitalisation. The AKI group had higher levels of urinary NAG, α1-microglobulin, ß2-microglobulin, urinary microalbumin, urinary transferrin and retinol-binding protein than the non-AKI group (P < 0.05). The MPP-related AKI prediction model consists of four indicators (serum immunoglobulin M [IgM], C-reactive protein [CRP], urine NAG and sputum plug presence) and a dynamic nomogram. CONCLUSION: Urinary kidney injury markers are often elevated in children with MPP; urinary NAG is the marker most likely to be elevated, and it is especially evident in severe cases. The nomogram of the prediction model, comprising serum IgM, CRP, urinary NAG and sputum plug presence, can predict the probability of AKI in children with MPP.
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Injúria Renal Aguda , Biomarcadores , Pneumonia por Mycoplasma , Humanos , Feminino , Masculino , Biomarcadores/urina , Pneumonia por Mycoplasma/complicações , Pneumonia por Mycoplasma/urina , Pneumonia por Mycoplasma/diagnóstico , Criança , Injúria Renal Aguda/urina , Injúria Renal Aguda/diagnóstico , Pré-Escolar , Nomogramas , Fatores de Risco , Valor Preditivo dos Testes , Modelos LogísticosRESUMO
Many cattle infected with Mycoplasma bovis remain healthy while others develop severe chronic respiratory disease. We hypothesized that inflammatory stimuli such as co-pathogens worsen disease outcomes in M. bovis-infected calves. Calves (n=24) were intrabronchially inoculated with M. bovis and either killed bacterial lysate, transient M. haemolytica infection, or saline. Caseonecrotic lesions developed in 7/7 animals given M. haemolytica and M. bovis compared to 2/8 given M. bovis with no inflammatory stimulus, and 6/9 animals given bacterial lysate and M. bovis (P=0.01). Animals receiving M. haemolytica and M. bovis had more caseonecrotic foci in lungs than those receiving M. bovis with no inflammatory stimulus (median = 21 vs 0; P = 0.01), with an intermediate response (median = 5) in animals given bacterial lysate. In addition to caseonecrotic foci, infected animals developed neutrophilic bronchiolitis that appeared to develop into caseonecrotic foci, peribronchiolar lymphocytic cuffs that were not associated with the other lesions, and 4 animals with bronchiolitis obliterans. The data showed that transient lung inflammation at the time of M. bovis infection provoked the development of caseonecrotic bronchopneumonia, and the severity of inflammation influenced the number of caseonecrotic foci that developed. In contrast, caseonecrotic lesions were few or absent in M. bovis-infected calves without a concurrent inflammatory stimulus. These studies provide insight into how caseonecrotic lesions develop within the lung of M. bovis-infected calves. This and other studies suggest that controlling co-pathogens and harmful inflammatory responses in animals infected with M. bovis could potentially minimize development of M. bovis caseonecrotic bronchopneumonia.