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.
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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.
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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: 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: 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
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.
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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.
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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 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.
Assuntos
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
Mycoplasma pneumoniae is a significant pathogen responsible for community-acquired pneumonia, predominantly affecting children and adolescents. Here, we devised a rapid method for M. pneumoniae that combined multiple cross displacement amplification (MCDA) with real-time fluorescence technology. A set of ten primers, which were specifically designed for M. pneumoniae detection, were employed in a real-time fluorescence MCDA reaction. Of these, one primer incorporated a restriction endonuclease recognition sequence, a fluorophore, and a quencher, facilitating real-time fluorescence detection. The real-time (RT)-MCDA reactions were monitored in a simple real-time fluorescence instrument and conducted under optimised conditions (64°C for 40 min). The detection limit of the M. pneumoniae RT-MCDA assay for genomic DNA extracted from M. pneumoniae culture was down to 43 fg/µl. This assay accurately identified M. pneumoniae strains without cross-reacting with other bacteria. To validate its practical application, we tested the M. pneumoniae RT-MCDA assay using genomic DNA extracted from clinical samples. The assay's detection capability proved comparable with real-time PCR, MCDA-based biosensor detection, and visual inspection under blue light. The entire process, including rapid DNA extraction and real-time MCDA detection, was completed within 1 h. Overall, the M. pneumoniae RT-MCDA assay reported here is a simple and effective diagnostic tool for rapid M. pneumoniae detection, which holds significant potential for point-of-care testing and in resource-limited regions.
Assuntos
DNA Bacteriano , Mycoplasma pneumoniae , Técnicas de Amplificação de Ácido Nucleico , Pneumonia por Mycoplasma , Sensibilidade e Especificidade , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/isolamento & purificação , Humanos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/microbiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Bacteriano/genética , Fluorescência , Técnicas de Diagnóstico Molecular/métodos , Primers do DNA/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Limite de DetecçãoRESUMO
BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) is a significant contributor to community-acquired pneumonia among children. Since 1968, when a strain of M. pneumoniae resistant to macrolide antibiotics was initially reported in Japan, macrolide-resistant M. pneumoniae (MRMP) has been documented in many countries worldwide, with varying incidence rates. MRMP infections lead to a poor response to macrolide antibiotics, frequently resulting in prolonged fever, extended antibiotic treatment, increased hospitalization, intensive care unit admissions, and a significantly higher proportion of patients receiving glucocorticoids or second-line antibiotics. Since 2000, the global incidence of MRMP has gradually increased, especially in East Asia, which has posed a serious challenge to the treatment of M. pneumoniae infections in children and attracted widespread attention from pediatricians. However, there is still no global consensus on the diagnosis and treatment of MRMP in children. METHODS: We organized 29 Chinese experts majoring in pediatric pulmonology and epidemiology to write the world's first consensus on the diagnosis and treatment of pediatric MRMP pneumonia, based on evidence collection. The evidence searches and reviews were conducted using electronic databases, including PubMed, Embase, Web of Science, CNKI, Medline, and the Cochrane Library. We used variations in terms for "macrolide-resistant", "Mycoplasma pneumoniae", "MP", "M. pneumoniae", "pneumonia", "MRMP", "lower respiratory tract infection", "Mycoplasma pneumoniae infection", "children", and "pediatric". RESULTS: Epidemiology, pathogenesis, clinical manifestations, early identification, laboratory examination, principles of antibiotic use, application of glucocorticoids and intravenous immunoglobulin, and precautions for bronchoscopy are highlighted. Early and rapid identification of gene mutations associated with MRMP is now available by polymerase chain reaction and fluorescent probe techniques in respiratory specimens. Although the resistance rate to macrolide remains high, it is fortunate that M. pneumoniae still maintains good in vitro sensitivity to second-line antibiotics such as tetracyclines and quinolones, making them an effective treatment option for patients with initial treatment failure caused by macrolide antibiotics. CONCLUSIONS: This consensus, based on international and national scientific evidence, provides scientific guidance for the diagnosis and treatment of MRMP in children. Further studies on tetracycline and quinolone drugs in children are urgently needed to evaluate their effects on the growth and development. Additionally, developing an antibiotic rotation treatment strategy is necessary to reduce the prevalence of MRMP strains.
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Antibacterianos , Farmacorresistência Bacteriana , Macrolídeos , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Pneumonia por Mycoplasma/tratamento farmacológico , Pneumonia por Mycoplasma/diagnóstico , Macrolídeos/uso terapêutico , Criança , Antibacterianos/uso terapêutico , Mycoplasma pneumoniae/efeitos dos fármacos , Masculino , Feminino , Pré-Escolar , Consenso , Infecções Comunitárias Adquiridas/tratamento farmacológico , Infecções Comunitárias Adquiridas/diagnóstico , Infecções Comunitárias Adquiridas/microbiologiaRESUMO
Since November 2023, the absolute number of attendances at emergency departments for pneumonia among children aged 5-14 years in England have been above expected levels for the time of year. This increased signal peaked during March 2024 but then persisted into early summer 2024 despite decreases in prevalence of seasonal respiratory pathogens. Record linkage between emergency department and laboratory databases points to this unusual activity being driven largely by Mycoplasma pneumoniae.
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Serviço Hospitalar de Emergência , Mycoplasma pneumoniae , Pneumonia , Humanos , Criança , Inglaterra/epidemiologia , Pré-Escolar , Adolescente , Incidência , Pneumonia/epidemiologia , Masculino , Feminino , Mycoplasma pneumoniae/isolamento & purificação , Serviço Hospitalar de Emergência/estatística & dados numéricos , Prevalência , Pneumonia por Mycoplasma/epidemiologia , Pneumonia por Mycoplasma/diagnóstico , Estações do Ano , Vigilância da PopulaçãoRESUMO
BACKGROUND: Pneumonia due to typical bacterial, atypical bacterial and viral pathogens can be difficult to clinically differentiate. Host response-based diagnostics are emerging as a complementary diagnostic strategy to pathogen detection. METHODS: We used murine models of typical bacterial, atypical bacterial and viral pneumonia to develop diagnostic signatures and understand the host's response to these types of infections. Mice were intranasally inoculated with Streptococcus pneumoniae, Mycoplasma pneumoniae, influenza or saline as a control. Peripheral blood gene expression analysis was performed at multiple time points. Differentially expressed genes were used to perform gene set enrichment analysis and generate diagnostic signatures. These murine-derived signatures were externally validated in silico using human gene expression data. The response to S. pneumoniae was the most rapid and robust. RESULTS: Mice infected with M. pneumoniae had a delayed response more similar to influenza-infected animals. Diagnostic signatures for the three types of infection had 0.94-1.00 area under the receiver operator curve (auROC). Validation in five human gene expression datasets revealed auROC of 0.82-0.96. DISCUSSION: This study identified discrete host responses to typical bacterial, atypical bacterial and viral aetiologies of pneumonia in mice. These signatures validated well in humans, highlighting the conserved nature of the host response to these pathogen classes.
Assuntos
Modelos Animais de Doenças , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Streptococcus pneumoniae , Animais , Humanos , Camundongos , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificação , Pneumonia por Mycoplasma/diagnóstico , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/isolamento & purificação , Feminino , Pneumonia Pneumocócica/microbiologia , Infecções por Orthomyxoviridae/imunologia , Curva ROC , Perfilação da Expressão Gênica , Pneumonia Viral/diagnóstico , Pneumonia Viral/imunologia , Camundongos Endogâmicos C57BL , Pneumonia Bacteriana/microbiologia , Pneumonia Bacteriana/diagnóstico , Interações Hospedeiro-PatógenoRESUMO
Mycoplasma pneumoniae (MP) is the cause of Mycoplasma pneumoniae pneumonia (MPP) in children and adolescents, with the clinical manifestations highlighted by intermittent irritating cough, accompanied by headache, fever and muscle pain. This paper aimed to study the research status and focal points in MP infection, especially the common laboratory diagnostic methods and clinical treatment of Mycoplasma pneumoniae. Laboratory diagnostic methods include molecular assay, serological antibody detection, rapid antigen detection and isolation and culture. Polymerase chain reaction (PCR) is the gold standard with high sensitivity and specificity. The serological antibody can detect various immune antibodies qualitatively or quantitatively in serum. Rapid antigen can be detected faster, with no equipment environment requirements, which can be used for the early diagnosis of MP infection. While the culture growth cycle is long and insensitive, not recommended for routine diagnosis. Macrolides were the preferred drug for children with MPP, while the drug resistance rate was rising in China. Tetracycline can be substituted but was not recommended for children under 8 years of age, quinolone drugs are not necessary, severe MPP can be combined with glucocorticoids, involving the nervous or immune system can choose gamma globulin. Other treatments for MPP including symptomatic treatment which can alleviate symptoms, improve lung function and improve prognosis. A safe and effective vaccine needed to be developed which can provide protective immunity to children and will reduce the incidence of MPP.
Assuntos
Antibacterianos , Mycoplasma pneumoniae , Pneumonia por Mycoplasma , Humanos , Criança , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/tratamento farmacológico , Mycoplasma pneumoniae/isolamento & purificação , Mycoplasma pneumoniae/imunologia , Antibacterianos/uso terapêutico , Adolescente , Pré-Escolar , Reação em Cadeia da Polimerase , Anticorpos Antibacterianos/sangue , Macrolídeos/uso terapêutico , Antígenos de Bactérias/imunologiaRESUMO
To determine the clinical indicators predictive of refractory Mycoplasma pneumoniae pneumonia (RMPP) in children and develop a robust predictive model to aid in early identification and management. A retrospective cohort study was conducted on 338 children diagnosed with RMPP out of a total of 1500 cases of Mycoplasma pneumoniae at a single tertiary hospital from May 2021 to November 2023. Clinical and demographic data analyzed included age, gender, parents' educational level, household income, body mass index, allergic constitution, and laboratory findings such as white blood cell count, neutrophil and lymphocyte counts, platelet count, and levels of C-reactive protein (CRP), D-dimer, and procalcitonin. Univariate and multivariate logistic regression analyses were performed to identify significant predictors of RMPP, and a predictive model was developed. Among the RMPP cohort, 52.4% were female, with a mean age of 6.07â ±â 2.78 years. Multivariate analysis identified several significant predictors of poor prognosis, including higher body mass index, longer duration of fever, elevated white blood cell count, neutrophil count, C-reactive protein levels, and increased neutrophil to lymphocyte ratio and platelet to lymphocyte ratio. The model demonstrated outstanding diagnostic performance, with an area under the receiver operating characteristic curve of 0.963 (95% confidence interval: 0.946-0.981). Our study identifies key clinical indicators with significant diagnostic accuracy for predicting RMPP in children. The predictive model established offers a valuable tool for clinicians, potentially improving RMPP outcomes through timely intervention.
Assuntos
Pneumonia por Mycoplasma , Humanos , Feminino , Masculino , Estudos Retrospectivos , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/sangue , Criança , Pré-Escolar , Proteína C-Reativa/análise , Prognóstico , Curva ROC , Contagem de Leucócitos , Mycoplasma pneumoniaeRESUMO
Objective: To explore the clinical characteristics and predictive factors for plastic bronchitis (PB) in children with severe Mycoplasma pneumoniae pneumonia (SMPP). Methods: A retrospective cohort enrolled children with a clinical diagnosis of SMPP who were treated at the Department of Respiratory Medicine of Tianjin Children's Hospital Machang District from January 1, 2018, to October 31, 2023. According to the bronchoscopy and pathological examination results, the patients were divided into 142 cases in the PB group and 274 cases in the non-PB group. The clinical manifestations, laboratory data, imaging findings, and treatments were analyzed.Mann-Whitney U test and Chi-square test were used to analyze the differences between the two groups, and multivariate Logistic regression was used to analyze the risk factors. The receiver operating characteristic (ROC) curve was used to explore the predictive value of PB in SMPP. Results: Among 416 SMPP children, there were 197 males and 219 females; PB group 142 cases, non-PB group 274 cases, the age of disease onset was (6.9±2.9) years and (6.6±2.8) years in the PB group and the non-PB group respectively. The incidence of wheezing symptoms, hypoxemia, heat peak >40 â, the duration of fever, neutrophil-lymphocyte ratio, mean platelet volume, C-reactive protein, procalcitonin, interleukin-6, alanine transaminase, aspartate aminotransferase and ferritin were higher in the PB group (16 cases (11.3%) vs. 15 cases (5.5%), 14 cases (9.9%) vs. 12 cases (4.4%), 57 cases (40.1%) vs. 67 cases (24.5%), 10 (8, 12) vs. 9 (8, 12) d, 6.1 (4.1, 13.1)×109 vs. 5.0 (3.7, 6.8)×109/L, 10.2 (9.6, 10.8) vs. 9.4 (8.9, 10.1) fl, 33.4 (16.0, 67.5) vs. 23.0 (10.4, 56.1) mg/L, 0.24 (0.12, 0.48) vs. 0.16 (0.09, 0.31) µg/L, 39.9 (25.1, 81.4) vs. 31.3 (18.3, 59.3) ng/L, 16.0 (12.0, 29.0) vs. 14.0 (10.0, 24.3) U/L, 38.5 (28.0, 52.5) vs. 33.0 (25.0, 44.0) U/L, 233 (136, 488) vs. 156 (110, 293) µg/L, χ2=4.55, 4.79, 11.00, Z=2.25, 4.00, 6.64, 2.76, 2.98, 3.09, 2.22, 2.62, 4.18, all P<0.05). Multivariate Logistic regression analysis showed that the dyspnea (OR=2.97, 95%CI 1.35-6.55, P=0.007), the diminution of respiration (OR=2.40, 95%CI 1.27-4.52, P=0.006), neutrophil-lymphocyte ratio (NLR) (OR=2.07, 95%CI 1.71-2.51, P<0.001), lactate dehydrogenase (LDH) (OR=1.01, 95%CI 1.00-1.01, P<0.001), mean platelet volume/platelet count (MPV/PLT) (OR=1.39, 95%CI 1.13-1.71, P=0.002), pleural effusion (OR=2.23, 95%CI 1.21-4.13, P=0.011),≥2/3 lobe consolidation (OR=1.84, 95%CI 1.04-3.00, P=0.039) and atelectasis (OR=1.98, 95%CI 1.02-3.48, P=0.044) were independent predictors of PB in children with SMPP. ROC curve analysis showed that the cut-off values for NLR, LDH and MPV/PLT in the diagnosis of PB were 2.79 (sensitivity 0.89, specificity 0.69, area under the curve (AUC)=0.86, P<0.001), 474 U/L (sensitivity 0.63, specificity 0.65, AUC=0.70, P=0.003) and 0.04 (sensitivity 0.75, specificity 0.53, AUC=0.68, P=0.005) respectively. Children in the PB group had longer hospital stays and corticosteroid treatment course than those in the non-PB group, the proportion of children in the PB group who received bronchoscopy treatment twice or more was higher (9 (8, 12) vs. 8 (6, 10) d, 7 (5, 8) vs. 6 (5, 7) d, 128 cases (90.1%) vs. 218 cases (79.6%), 106 cases (74.7%) vs. 54 cases (19.7%), Z=6.70, 5.06, χ2=7.48, 119.27, all P<0.05). Conclusions: The dyspnea, respiration diminution, NLR level elevation (>2.79) and pleural effusion were predictive factors for PB in children with SMPP. This provides a basis for the early identification of PB in children with SMPP.
Assuntos
Bronquite , Pneumonia por Mycoplasma , Humanos , Masculino , Feminino , Pneumonia por Mycoplasma/diagnóstico , Estudos Retrospectivos , Criança , Bronquite/diagnóstico , Pré-Escolar , Fatores de Risco , Proteína C-Reativa/análise , Proteína C-Reativa/metabolismo , Mycoplasma pneumoniae , Pró-Calcitonina/sangue , Curva ROC , Modelos Logísticos , Volume Plaquetário Médio , Ferritinas/sangue , Febre , Índice de Gravidade de DoençaRESUMO
Mycoplasma pneumoniae pneumonia (MPP) is a common respiratory tract infection disease in children. To date, there have been few studies on the relationship between cytological changes in bronchoalveolar lavage fluid (BALF) and clinical features. The objective of this study is to investigate the correlation between changes in the proportion of cell classifications in BALF and the clinical features in children with severe MPP (SMPP). In total, the study included 64 children with SMPP requiring bronchoalveolar lavage who were admitted to our hospital between March and September 2022 (study group) and 11 children with bronchial foreign bodies without co-infection (control group), who were admitted during the same period. The proportion of cell classifications in BALF was determined by microscopic examination after performing Wright-Giemsa staining. Patients were grouped according to different clinical characteristics, and between-group comparisons were made regarding the variations in the proportion of cell classifications in BALF. The levels of blood routine neutrophil percentage (GRA%), C-reactive protein, D-dimer and lactate dehydrogenase in the study group were higher than those in the control group (P < 0.05). There were differences in the GRA% and macrophage percentage in the BALF between the two groups (P < 0.05). The GRA% and blood lymphocyte percentage were associated with pleural effusion. Multiple indicators correlated with extrapulmonary manifestations (P < 0.05). Moreover, the percentage of lymphocytes in the BALF correlated with pleural effusion, extrapulmonary manifestations and refractory MPP (RMPP) (P < 0.05). Logistic regression showed that BALF lymphocytes were protective factors for RMPP, while serum amyloid A and extrapulmonary manifestations were risk factors (P < 0.05). CONCLUSION: The BALF of children with SMPP is predominantly neutrophilic. A lower percentage of lymphocytes is related to a higher incidence of pleural effusion, extrapulmonary manifestations and progression to RMPP, as well as a longer length of hospitalisation. WHAT IS KNOWN: ⢠Mycoplasma pneumonia in children is relatively common in clinical practice. Bronchoalveolar lavage (BAL) is a routine clinical procedure. WHAT IS NEW: However, there are relatively few studies focusing on the cytomorphological analysis of cells in BAL fluid.