RESUMEN

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.

Asunto(s)

Sistemas CRISPR-Cas , Mycoplasma pneumoniae , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Humanos , Sistemas CRISPR-Cas/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología

RESUMEN

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.

Asunto(s)

Agammaglobulinemia , Enfermedades Genéticas Ligadas al Cromosoma X , Neumonía por Mycoplasma , Humanos , Agammaglobulinemia/complicaciones , Agammaglobulinemia/genética , Enfermedades Genéticas Ligadas al Cromosoma X/genética , Enfermedades Genéticas Ligadas al Cromosoma X/complicaciones , Masculino , Neumonía por Mycoplasma/complicaciones , Neumonía por Mycoplasma/microbiología , Agammaglobulinemia Tirosina Quinasa/genética , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Adulto , Secuenciación de Nucleótidos de Alto Rendimiento

RESUMEN

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.

Asunto(s)

MicroARNs , Neumonía por Mycoplasma , Humanos , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/sangre , Masculino , Femenino , MicroARNs/sangre , Niño , Preescolar , Biomarcadores/sangre , Mycoplasma pneumoniae/genética

RESUMEN

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.

Asunto(s)

Sistemas CRISPR-Cas , Técnicas de Diagnóstico Molecular , Mycoplasma pneumoniae , Técnicas de Amplificación de Ácido Nucleico , Neumonía por Mycoplasma , Sensibilidad y Especificidad , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Humanos , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología , Técnicas de Diagnóstico Molecular/métodos , Niño , Límite de Detección

RESUMEN

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.

Asunto(s)

ADN Bacteriano , Mycoplasma pneumoniae , Técnicas de Amplificación de Ácido Nucleico , Neumonía por Mycoplasma , Sensibilidad y Especificidad , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Humanos , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología , Técnicas de Amplificación de Ácido Nucleico/métodos , ADN Bacteriano/genética , Fluorescencia , Técnicas de Diagnóstico Molecular/métodos , Cartilla de ADN/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Límite de Detección

RESUMEN

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.

Asunto(s)

Modelos Animales de Enfermedad , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Streptococcus pneumoniae , Animales , Humanos , Ratones , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/aislamiento & purificación , Neumonía por Mycoplasma/diagnóstico , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Femenino , Neumonía Neumocócica/microbiología , Infecciones por Orthomyxoviridae/inmunología , Curva ROC , Perfilación de la Expresión Génica , Neumonía Viral/diagnóstico , Neumonía Viral/inmunología , Ratones Endogámicos C57BL , Neumonía Bacteriana/microbiología , Neumonía Bacteriana/diagnóstico , Interacciones Huésped-Patógeno

RESUMEN

BACKGROUNDS: Mycoplasma pneumoniae (M. pneumoniae) is a common pathogen causing respiratory diseases in children. This study aimed to characterize epidemiological and disease severity shifts of M. pneumoniae: infections in Guangzhou, China during and after the coronavirus disease 2019 (COVID-19) pandemic. METHODS: Throat swab samples were obtained from 5405 hospitalized patients with symptoms of acute respiratory infections to detect M. pneumoniae. Differences in epidemiological and clinical characteristics of M. pneumoniae: infections were investigated during 2020-2022 and after COVID-19 pandemic (2023). RESULTS: M. pneumoniae were detected in 849 (15.6%, 849/5405) patients. The highest annual positive rate was 29.4% (754/2570) in 2023, followed by 5.3% (72/1367) in 2022, 1.2% (12/1015) in 2021, and 2.0% (11/553) in 2020, with significantly increasing annual prevalence from 2020 to 2023. M. pneumoniae incidence peaked between July and December post-COVID-19 pandemic in 2023, with the highest monthly positive rate (56.4%, 165/293). Clinical characteristics and outcomes of patients with M. pneumoniae did not vary between periods during and after COVID-19 pandemic except that patients with M. pneumoniae post-COVID-19 pandemic were more likely to develop fever. Patients with severe M. pneumoniae pneumonia (SMPP) were more likely to develop respiratory complications, myocardial damage, and gastrointestinal dysfunction than those with non-SMPP. Patients with SMPP had lower lymphocytes, CD3+ T cells, CD4+ T cells, CD8+ T cells, B cells, and higher IL-4, IL-6, IL-10 levels than those with non-SMPP. Bronchoalveolar lavage fluid specimens from infected patients were obtained to identify macrolide resistance mutations. Macrolide-resistant M. pneumoniae (MRMP) proportion in 2023 was 91.1% (215/236). CONCLUSION: Outbreaks of M. pneumoniae: occurred in Guangzhou, China in 2023 upon Non-pharmaceutical interventions easing. Despite the increasing incidence of M. pneumoniae, the disease severity remained similar during and after the COVID-19 pandemic.

Asunto(s)

COVID-19 , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , China/epidemiología , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/microbiología , COVID-19/epidemiología , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Masculino , Femenino , Niño , Adulto , Adolescente , Persona de Mediana Edad , Preescolar , Adulto Joven , Brotes de Enfermedades , SARS-CoV-2/genética , Lactante , Anciano , Incidencia , Prevalencia , Pandemias

RESUMEN

OBJECTIVE: This study aimed to investigate the pathogen epidemiology of community-acquired pneumonia (CAP) among children in Southwest China before, during and after the COVID-19 non-pharmaceutical interventions (NPIs). METHODS: Pathogen data of hospitalised children with CAP, including multiple direct immunofluorescence test for seven viruses, bacterial culture and polymerase chain reaction (PCR) for Mycoplasma pneumoniae, were analysed across three phases: Phase I (pre-NPIs: 1 January 2019 to 31 December 2019), Phase II (NPI period: 1 January 2020 to 31 December 2020) and Phase III (post-NPIs: 1 January 2023 to 31 December 2023). RESULTS: A total of 7533 cases were enrolled, including 2444, 1642 and 3447 individuals in Phases I, II and III, respectively. M. pneumoniae predominated in Phases I and III (23.4% and 35.5%, respectively). In Phase II, respiratory syncytial virus (RSV) emerged as the primary pathogen (20.3%), whereas detection rates of influenza A virus (Flu A) and M. pneumoniae were at a low level (1.8% and 9.6%, respectively). In Phase III, both Flu A (15.8%) and M. pneumoniae epidemic rebounded, whereas RSV detection rate returned to Phase I level, and detection rates of Streptococcus pneumoniae and Haemophilus influenzae decreased significantly compared to those in Phase I. Detection rates of adenovirus and parainfluenza virus type 3 decreased phase by phase. Age-stratified analysis and monthly variations supported the above findings. Seasonal patterns of multiple pathogens were disrupted during Phases II and III. CONCLUSIONS: COVID-19 NPIs exhibited a distinct impact on CAP pathogen epidemic among children, with post-NPIs increases observed in M. pneumoniae and Flu A prevalence. Continuous pathogen monitoring is crucial for effective prevention and control of paediatric CAP.

Asunto(s)

COVID-19 , Infecciones Comunitarias Adquiridas , Humanos , Infecciones Comunitarias Adquiridas/epidemiología , Infecciones Comunitarias Adquiridas/microbiología , Infecciones Comunitarias Adquiridas/virología , China/epidemiología , COVID-19/epidemiología , Estudios Transversales , Preescolar , Femenino , Masculino , Niño , Lactante , SARS-CoV-2/aislamiento & purificación , SARS-CoV-2/genética , Mycoplasma pneumoniae/aislamiento & purificación , Mycoplasma pneumoniae/genética , Adolescente , Neumonía/epidemiología , Neumonía/microbiología , Neumonía/virología

RESUMEN

PURPOSE: To investigate macrolide-resistant Mycobacterium pneumoniae (MRMP) pneumonia in children and construct a logistic regression model for mutations in the Mycoplasma pneumoniae drug-resistant gene. METHODS: Clinical data of 281 children were analyzed. Sequencing confirmed a mutation at the A2063G locus of the 23 S rRNA gene in 227 children (A2063G group); 54 children showed no mutations (non-MRMP [NMRMP] group). We compared clinical features, laboratory tests, imaging, and bronchoscopy results and constructed a multifactorial logistic regression model to analyze risk and protective factors. RESULTS: The A2063G group had longer durations of fever and hospitalization before admission, a higher proportion of treatment with sodium methylprednisolone succinate (MPS)/dexamethasone, longer time to discontinue hormones, and higher probability of combined infections. Monocyte percentage was significantly higher in the A2063G group. Imaging suggested a higher incidence of infections in the right lung compared to both lungs. Univariate analysis revealed fever duration before admission, hormone dose and duration, monocyte percentage, and mixed infections as risk factors for Mycoplasma pneumoniae infection with the A2063G mutation. The logistic regression model showed that mixed infections were an independent risk factor for the A2063G locus mutation, whereas hormone dose was a protective factor. CONCLUSION: A prevalence of macrolide resistance of 80.8% among children was observed in the region. Logistic regression analysis revealed that co-infection with other respiratory pathogens is an independent risk factor for the development of resistance genes, while the use of hormone dosage acts as a protective factor.

Asunto(s)

Antibacterianos , Farmacorresistencia Bacteriana , Macrólidos , Mycoplasma pneumoniae , Neumonía por Mycoplasma , ARN Ribosómico 23S , Humanos , Neumonía por Mycoplasma/microbiología , Neumonía por Mycoplasma/tratamiento farmacológico , Neumonía por Mycoplasma/epidemiología , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/efectos de los fármacos , Macrólidos/farmacología , Macrólidos/uso terapéutico , Femenino , Masculino , Farmacorresistencia Bacteriana/genética , Preescolar , Niño , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , ARN Ribosómico 23S/genética , Modelos Logísticos , Lactante , Mutación , Factores de Riesgo , Estudios Retrospectivos

Asunto(s)

Técnicas de Diagnóstico Molecular , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Sensibilidad y Especificidad , Humanos , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología , Técnicas de Diagnóstico Molecular/métodos , Niño , Preescolar , Adulto , Adolescente , Masculino , Sistema Respiratorio/microbiología , Femenino , Persona de Mediana Edad

RESUMEN

AIMS: A severe lockdown occurred in Wuhan during the COVID-19 pandemic, followed by a remission phase in the pandemic's aftermath. This study analyzed the bacterial and fungal profiles of respiratory pathogens in patients hospitalized with non-COVID-19 lower respiratory tract infections (LRTIs) during this period to determine the pathogen profile distributions in different age groups and hospital departments in Wuhan. METHODS AND RESULTS: We collected reports of pathogen testing in the medical records of patients hospitalized with non-COVID-19 LRTI between 2019 and 2021. These cases were tested for bacterial and fungal pathogens using 16S and internal transcribed spacer sequencing methods on bronchoalveolar lavage fluid samples. The study included 1368 cases. The bacteria most commonly identified were Streptococcus pneumoniae (12.50%) and Mycoplasma pneumoniae (8.33%). The most commonly identified fungi were Aspergillus fumigatus (2.49%) and Pneumocystis jirovecii (1.75%). Compared to 2019, the S. pneumoniae detection rates increased significantly in 2021, and those of M. pneumoniae decreased. Streptococcus pneumoniae was detected mainly in children. The detection rates of almost all fungi were greater in the respiratory Intensive Care Unit compared to respiratory medicine. Streptococcus pneumoniae and M. pneumoniae were detected more frequently in the pediatric department. CONCLUSIONS: Before and after the COVID-19 outbreak, a change in the common pathogen spectrum was detected in patients with non-COVID-19 in Wuhan, with the greatest change occurring among children. The major pathogens varied by the patient's age and the hospital department.

Asunto(s)

COVID-19 , Hospitalización , Infecciones del Sistema Respiratorio , Humanos , China/epidemiología , Infecciones del Sistema Respiratorio/microbiología , Infecciones del Sistema Respiratorio/virología , Infecciones del Sistema Respiratorio/epidemiología , Persona de Mediana Edad , Niño , Masculino , Adulto , Femenino , Preescolar , Adolescente , Anciano , Lactante , COVID-19/epidemiología , Hongos/aislamiento & purificación , Hongos/genética , Hongos/clasificación , Adulto Joven , Streptococcus pneumoniae/aislamiento & purificación , Streptococcus pneumoniae/genética , Bacterias/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , Mycoplasma pneumoniae/aislamiento & purificación , Mycoplasma pneumoniae/genética , Líquido del Lavado Bronquioalveolar/microbiología , Líquido del Lavado Bronquioalveolar/virología

RESUMEN

Before the COVID-19 pandemic, Mycoplasma pneumoniae infections emerged during spring to summer yearly in Taiwan, but infections were few during the pandemic. M. pneumoniae macrolide resistance soared to 85.7% in 2020 but declined to 0% during 2022-2023. Continued molecular surveillance is necessary to monitor trends in macrolide-resistant M. pneumoniae.

Asunto(s)

Antibacterianos , COVID-19 , Farmacorresistencia Bacteriana , Macrólidos , Mycoplasma pneumoniae , Neumonía por Mycoplasma , SARS-CoV-2 , Humanos , Taiwán/epidemiología , Macrólidos/farmacología , Macrólidos/uso terapéutico , Mycoplasma pneumoniae/efectos de los fármacos , Mycoplasma pneumoniae/genética , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/tratamiento farmacológico , Neumonía por Mycoplasma/microbiología , COVID-19/epidemiología , Niño , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Preescolar , Pandemias , Masculino , Femenino , Lactante , Adolescente , Pruebas de Sensibilidad Microbiana

RESUMEN

Mycoplasma pneumoniae is a common cause of community-acquired pneumonia in children and young adults. It is responsible of a broad array of extrapulmonary manifestations, the most severe affecting the central nervous system. We report a challenging diagnosis of macrolide-resistant M. pneumoniae-induced Bickerstaff encephalitis in a 16-year-old man.

Asunto(s)

Encefalitis , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , Mycoplasma pneumoniae/aislamiento & purificación , Mycoplasma pneumoniae/genética , Adolescente , Masculino , Encefalitis/microbiología , Encefalitis/diagnóstico , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/microbiología , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/tratamiento farmacológico , Brotes de Enfermedades , Antibacterianos/uso terapéutico , Farmacorresistencia Bacteriana , Infecciones Comunitarias Adquiridas/microbiología , Infecciones Comunitarias Adquiridas/diagnóstico , Infecciones Comunitarias Adquiridas/epidemiología , Macrólidos/uso terapéutico

RESUMEN

To analyze the infection and drug-resistant gene 23S rRNA mutations of mycoplasma pneumoniae (Mp) in hospitalized children aged 0-17 in Ningbo City from 2019 to 2023. Throat swabs were collected from hospitalized children with respiratory tract infections in Ningbo University Affiliated Women and Children's Hospital from 2019 to 2023. They were subjected to real-time fluorescence quantitative polymerase chain reaction detection to analyze Mp infection and drug-resistant gene (23S rRNA) mutations. Intergroup comparisons were made by the Chi-square test or Fisher's exact probability method. A total of 18 968 hospitalized children were included, with a total positive rate of 30.37% (5 760/18 968). The total positive rate of drug-resistant gene mutations was 82.45% (4 749/5 760). The positive rate of Mp in male children was 29.26%, which was lower than that in female children (31.67%, χ2=12.948, P<0.001). The positive rate of Mp drug-resistant gene mutations in male children was 82.52%, which was higher than that in female children(82.37%, χ2=0.021, P=0.885). The positive rates of Mp increased with age (χ2=1 722.21, P<0.001). The positive rates of Mp drug-resistant gene mutations also increased with age (χ2=13.152, P<0.001). In the four seasons, the total positive rate of Mp in summer and autumn was significantly higher than that in winter and spring (χ2=1 085.149, P<0.001). Among them, the Mp positive rates in the summer and autumn of 2019 were as high as 38.26% and 34.49%, while in the summer and autumn of 2020, the Mp positive rates were 2.55% and 1.65%, respectively, which were the lowest in previous years. In the summer and autumn of 2023, the Mp positive rates increased to 47.22% and 51.06%. There was no statistically significant difference in the detection rate of Mp drug-resistant gene mutations among the four seasons. In Conclusion, Mp infection was more prevalent in the summer and autumn in Ningbo city and females and children aged 7-17 were more susceptible. The epidemic of Mp infection in Ningbo occurred in the summer of 2019. After the COVID-19 pandemic in 2020, the positive rate of Mp rapidly decreased and later remained in a low incidence state. After the lifting of restrictive prevention and control measures in 2023, the Mp positive rate returned to an epidemic state. The positive rate of Mp drug-resistant gene (23S rRNA) mutations was relatively high.

Asunto(s)

Farmacorresistencia Bacteriana , Mutación , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , Niño , Lactante , Preescolar , Femenino , Masculino , Mycoplasma pneumoniae/genética , Adolescente , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/microbiología , Farmacorresistencia Bacteriana/genética , ARN Ribosómico 23S/genética , Infecciones del Sistema Respiratorio/microbiología , Infecciones del Sistema Respiratorio/epidemiología , Recién Nacido , China/epidemiología , Antibacterianos/farmacología , Antibacterianos/uso terapéutico

RESUMEN

BACKGROUND: The global prospective surveillance data showed the re-emergence of mycoplasma pneumoniae pneumonia (MPP) in Europe and Asia after the coronavirus disease 2019 pandemic. We sought to observe the effect of macrolide antibiotics in the treatment of MPP carrying a macrolide-resistant mutation gene and the potential of targeted next-generation sequencing (tNGS) as a front-line diagnostic in MPP patients. METHODS: The baseline characteristics of 91 children with MPP hospitalized from January to October 2023 were retrospectively analyzed. They were divided into two groups according to whether carrying the macrolide-resistant mutation or not. The logistic and linear regression analyses were used to determine whether the mutation was a standalone predictive predictor of the duration of fever and hospital length of stay. RESULTS: First, no patients had a fever for ≥ 7 days after macrolide treatment. But length of stay and hormone concentration were significantly different between the two groups (P < 0.05). There were also no statistical association between the mutation and the duration of fever and hospital length of stay. CONCLUSION: Macrolides can be administered to MPP children carrying a macrolide-resistant mutation. tNGS can be seen as a front-line diagnostic in MPP.

Asunto(s)

Antibacterianos , Farmacorresistencia Bacteriana , Macrólidos , Mutación , Mycoplasma pneumoniae , Neumonía por Mycoplasma , ARN Ribosómico 23S , Humanos , Neumonía por Mycoplasma/tratamiento farmacológico , Neumonía por Mycoplasma/microbiología , Macrólidos/uso terapéutico , Macrólidos/farmacología , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/efectos de los fármacos , Femenino , Masculino , Antibacterianos/uso terapéutico , Antibacterianos/farmacología , Preescolar , Niño , Farmacorresistencia Bacteriana/genética , Estudios Retrospectivos , ARN Ribosómico 23S/genética , Lactante , Tiempo de Internación , Resultado del Tratamiento , Secuenciación de Nucleótidos de Alto Rendimiento

RESUMEN

To analyze the characteristics of Mycoplasma pneumoniae as well as macrolide antibiotic resistance through whole-genome sequencing and comparative genomics. Thirteen clinical strains isolated from 2003 to 2019 were selected, 10 of which were resistant to erythromycin (MIC >64 µg/mL), including 8 P1-type I and 2 P1-type II. Three were sensitive (<1 µg/mL) and P1-type II. One resistant strain had an A→G point mutation at position 2064 in region V of the 23S rRNA, the others had it at position 2063, while the three sensitive strains had no mutation here. Genome assembly and comparative genome analysis revealed a high level of genome consistency within the P1 type, and the primary differences in genome sequences concentrated in the region encoding the P1 protein. In P1-type II strains, three specific gene mutations were identified: C162A and A430G in L4 gene and T1112G mutation in the CARDS gene. Clinical information showed seven cases were diagnosed with severe pneumonia, all of which were infected with drug-resistant strains. Notably, BS610A4 and CYM219A1 exhibited a gene multi-copy phenomenon and shared a conserved functional domain with the DUF31 protein family. Clinically, the patients had severe refractory pneumonia, with pleural effusion, necessitating treatment with glucocorticoids and bronchoalveolar lavage. The primary variations between strains occur among different P1-types, while there is a high level of genomic consistency within P1-types. Three mutation loci associated with specific types were identified, and no specific genetic alterations directly related to clinical presentation were observed.IMPORTANCEMycoplasma pneumoniae is an important pathogen of community-acquired pneumonia, and macrolide resistance brings difficulties to clinical treatment. We analyzed the characteristics of M. pneumoniae as well as macrolide antibiotic resistance through whole-genome sequencing and comparative genomics. The work addressed primary variations between strains that occur among different P1-types, while there is a high level of genomic consistency within P1-types. In P1-type II strains, three specific gene mutations were identified: C162A and A430G in L4 gene and T1112G mutation in the CARDS gene. All the strains isolated from severe pneumonia cases were drug-resistant, two of which exhibited a gene multi-copy phenomenon, sharing a conserved functional domain with the DUF31 protein family. Three mutation loci associated with specific types were identified, and no specific genetic alterations directly related to clinical presentation were observed.

Asunto(s)

Antibacterianos , Farmacorresistencia Bacteriana , Genoma Bacteriano , Pruebas de Sensibilidad Microbiana , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/efectos de los fármacos , Mycoplasma pneumoniae/clasificación , Mycoplasma pneumoniae/aislamiento & purificación , Humanos , Antibacterianos/farmacología , Genoma Bacteriano/genética , Neumonía por Mycoplasma/microbiología , Neumonía por Mycoplasma/tratamiento farmacológico , Farmacorresistencia Bacteriana/genética , Masculino , Femenino , Secuenciación Completa del Genoma , Persona de Mediana Edad , Macrólidos/farmacología , Adulto , Mutación , ARN Ribosómico 23S/genética , Genómica , Anciano , Eritromicina/farmacología

Asunto(s)

Coinfección , Absceso Pulmonar , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , Mycoplasma pneumoniae/genética , Coinfección/microbiología , Preescolar , Neumonía por Mycoplasma/complicaciones , Neumonía por Mycoplasma/microbiología , Absceso Pulmonar/microbiología , Absceso Pulmonar/tratamiento farmacológico , Masculino , Antibacterianos/uso terapéutico

RESUMEN

With the atypical rise of Mycoplasma pneumoniae infection (MPI) in 2023, prompt studies are needed to determine the current epidemic features and risk factors with emerging trends of MPI to furnish a framework for subsequent investigations. This multicentre, retrospective study was designed to analyse the epidemic patterns of MPI before and after the COVID-19 pandemic, as well as genotypes and the macrolide-resistance-associated mutations in MP sampled from paediatric patients in Southern China. Clinical data was collected from 1,33,674 patients admitted into investigational hospitals from 1 June 2017 to 30 November 2023. Metagenomic next-generation sequencing (mNGS) data were retrieved based on MP sequence positive samples from 299 paediatric patients for macrolide-resistance-associated mutations analysis. Pearson's chi-squared test was used to compare categorical variables between different time frames. The monthly average cases of paediatric common respiratory infection diseases increased without enhanced public health measures after the pandemic, especially for influenza, respiratory syncytial virus infection, and MPI. The contribution of MPI to pneumoniae was similar to that in the outbreak in 2019. Compared to mNGS data between 2019-2022 and 2023, the severity of MP did not grow stronger despite higher rates of macrolide-resistance hypervariable sites, including loci 2063 and 2064, were detected in childhood MP samples of 2023. Our findings indicated that ongoing surveillance is necessary to understand the impact of post pandemic on MP transmission disruption during epidemic season and the severity of clinical outcomes in different scenarios.

Asunto(s)

COVID-19 , Mycoplasma pneumoniae , Neumonía por Mycoplasma , Humanos , Neumonía por Mycoplasma/epidemiología , Neumonía por Mycoplasma/microbiología , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Mycoplasma pneumoniae/efectos de los fármacos , China/epidemiología , COVID-19/epidemiología , COVID-19/transmisión , Niño , Estudios Retrospectivos , Preescolar , Masculino , Femenino , Lactante , Macrólidos/farmacología , Farmacorresistencia Bacteriana , SARS-CoV-2/genética , SARS-CoV-2/aislamiento & purificación , Adolescente , Secuenciación de Nucleótidos de Alto Rendimiento , Antibacterianos/farmacología , Pandemias

RESUMEN

A concise and rapid detection method for Mycoplasma pneumoniae is urgently required due to its severe impact on human health. To meet such a need, this study proposed and constructed an innovative point-of-care testing (POCT) platform that consists of a hydrogen ion-selective loop-mediated isothermal amplification (H+-LAMP) sensor and an electrochemical detection device. The H+-LAMP sensor successfully integrated the working and reference electrodes and converted the H+ generated during the LAMP process into an electrochemical signal. High sensitivity and stability for pathogen detection were also achieved by treating the working electrode with an electrodeposited polyaniline solid contact layer and by using an ion-selective membrane. As a result, the sensor shows a sensitivity of 68.26 mV per pH, a response time of less than 2 s, and a potential drift of less than 5 mV within one hour, which well meets the urgent need. The results also demonstrated that the detection limit for Mycoplasma pneumoniae was lowered to 1 copy per µL, the nucleic acid extraction and detection process could be completed in 30 minutes, and the impact of interfering ions on the sensor was negligible. Validation with 20 clinical samples yielded satisfactory results. More importantly, the storage lifespan of such an electrochemical sensor is over seven days, which is a great advantage for on-site pathogen detection. Therefore, the hydrogen ion-selective sensor constructed in this investigation is particularly suitable as a core component for instant pathogen detection platforms.

Asunto(s)

Técnicas Electroquímicas , Límite de Detección , Mycoplasma pneumoniae , Técnicas de Amplificación de Ácido Nucleico , Mycoplasma pneumoniae/aislamiento & purificación , Mycoplasma pneumoniae/genética , Técnicas Electroquímicas/métodos , Técnicas Electroquímicas/instrumentación , Técnicas de Amplificación de Ácido Nucleico/métodos , Humanos , Hidrógeno/química , Neumonía por Mycoplasma/diagnóstico , Neumonía por Mycoplasma/microbiología , Técnicas Biosensibles/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/instrumentación , Electrodos

RESUMEN

To address the limitations of the CRISPR/Cas12f1 system in clinical diagnostics, which require the complex preparation of single-stranded DNA (ssDNA) or in vitro transcripts (RNA), we developed a fluorescent biosensor named PDTCTR (PAM-dependent dsDNA Target-activated Cas12f1 Trans Reporter). This innovative biosensor integrates Recombinase Polymerase Amplification (RPA) with the Cas12f_ge4.1 system, facilitating the direct detection of double-stranded DNA (dsDNA). PDTCTR represents a significant leap forward, exhibiting a detection sensitivity that is a hundredfold greater than the original Cas12f1 system. It demonstrates the capability to detect Mycoplasma pneumoniae (M. pneumoniae) and Hepatitis B virus (HBV) with excellent sensitivity of 10 copies per microliter (16.8 aM) and distinguishes single nucleotide variations (SNVs) with high precision, including the EGFR (L858R) mutations prevalent in non-small cell lung cancer (NSCLC). Clinical evaluations of PDTCTR have demonstrated its high sensitivity and specificity, with rates ranging from 93%-100% and 100%, respectively, highlighting its potential to revolutionize diagnostic approaches for infectious diseases and cancer-related SNVs.This research underscores the substantial advancements in CRISPR technology for clinical diagnostics and its promising future in early disease detection and personalized medicine.

Asunto(s)

Técnicas Biosensibles , Sistemas CRISPR-Cas , ARN Guía de Sistemas CRISPR-Cas , Técnicas Biosensibles/métodos , Humanos , ARN Guía de Sistemas CRISPR-Cas/genética , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/aislamiento & purificación , ADN/genética , ADN/química , Mycoplasma pneumoniae/genética , Mycoplasma pneumoniae/aislamiento & purificación , Proteínas Asociadas a CRISPR/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Proteínas Bacterianas/genética , Proteínas Bacterianas/química , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/diagnóstico , Endodesoxirribonucleasas/genética , Endodesoxirribonucleasas/química , Neumonía por Mycoplasma/diagnóstico