Mycoplasma pneumoniae infection outbreak in Guangzhou, China after COVID-19 pandemic.

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.

Mycoplasma pneumoniae epidemic in Denmark, October to December, 2023.

We report a surge of patients, especially children and adolescents, with respiratory disease caused by Mycoplasma pneumoniae in Denmark since October 2023. While the surge has reached an epidemic level, no impact on hospital capacity has been observed; only 14% (446/3,195) of cases, primarily adults, required hospitalisation. Macrolide resistance was detected in less than 2% of samples tested. Timely monitoring of hospitalisations linked to M. pneumoniae infections has been established to inform the healthcare system, decisionmakers and the public.

A case of anti-myelin oligodendrocyte glycoprotein (MOG)-immunoglobulin G (IgG) associated disorder (MOGAD) with clinical manifestations of acute disseminated encephalomyelitis: Secondary to mycoplasma pneumoniae infection.

Anti-myelin oligodendrocyte glycoprotein (MOG)-immunoglobulin G (IgG) associated disorder (MOGAD) is an immune-mediated central nervous system (CNS) inflammatory demyelinating disorder that has been widely recognized in recent years. It is distinct from multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD), which are separate disease spectrums. Here we report the case of a 5-year-old boy who was admitted for 3 days with fever, headache, and vomiting. Magnetic resonance imaging revealed abnormal hyperintensity in the left thalamus and positive serum IgM for M. pneumoniae. After treatment with azithromycin, the headache gradually disappeared, but paralysis and urinary retention occurred on the 6th day after admission. MRI re-examination showed that the original abnormal signal in the left thalamus was significantly weakened, but new abnormal signals appeared in the brain and cerebrospinal cord, and the serum MOG-IgG was positive. After treatment, the child has fully recovered and is still receiving follow-up care. We believe that this is a case of MOGAD in a child with a biphasic ADEM phenotype secondary to M. pneumoniae infection, which has potential value in elucidating the pathophysiology of MOGAD.

Epidemic characteristics of Mycoplasma pneumoniae infection: a retrospective analysis of a single center in Suzhou from 2014 to 2020.

Background: Mycoplasma pneumoniae (M. Pneumoniae) is a common pathogen of respiratory tract infections, but there is still a lack of detailed investigation on the large sample of M. Pneumoniae infection in the all age population. And patients with severe M. Pneumoniae pneumonia (SMPP) still have a certain risk of death. How to identify the clinical characteristics and population of patients with SMPP as soon as possible is still an urgent problem in clinical practice. Methods: Demographic characteristics, patient clinical information, and laboratory data of 81,131 patients with respiratory tract infections (RTIs) in the Affiliated Suzhou Hospital of Nanjing Medical University from 2014 to 2020 were retrospectively collected from all patient records. The serum particle agglutination (PA) test was used to determine M. Pneumoniae infection by detecting specific antibodies. The white blood cell count, the proportion of neutrophils and lymphocytes, C-reactive protein (CRP) and lactate dehydrogenase (LDH) levels between children and adults with SMPP were compared by Student's t-test; other clinical features were analyzed by χ2 test or Fisher's exact test. Results: A total of 81,131 patients with RTIs were included, and 21,582 (26.60%) M. Pneumoniae immunoglobulin M (IgM)-positive patients were detected. From 2014 to 2020, the annual proportions of M. Pneumoniae RTIs were 23.60%, 28.18%, 38.08%, 27.05%, 23.44%, 25.26%, and 18.33%, respectively. In terms of seasonal distribution, April-June and September-November were the peak seasons of M. Pneumoniae infection each year. Children and women have a high proportion of M. Pneumoniae infection. The peak age of M. Pneumoniae infection was between 4 and 14 years old. There were 301 cases of SMPP, including 281 children and 20 adults (8 cases of pregnant women). Children and pregnant women accounted for a high proportion of SMPP. Children with SMPP had more extrapulmonary symptoms, multilobar infiltrates, and increased CRP and LDH levels compared with adults. Conclusions: M. Pneumoniae infection has seasonal, sex, and age distribution trends. Children and pregnant women accounted for a high proportion of SMPP. Extrapulmonary symptoms, multilobar infiltrates, and increased CRP and LDH levels may be helpful to identify SMPP in children than in adults.

Assessment of variables associated with prolonged admission duration in children with Mycoplasma pneumoniae pneumonia.

INTRODUCTION: Macrolide-resistant Mycoplasma pneumoniae (MRMP) has become prevalent in children. This study investigated the clinical and laboratory variables of MRMP and macrolide-sensitive M. pneumoniae (MSMP) and identified factors associated with prolonged hospital admission in children. METHODS: A prospective multicenter study was conducted in 1063 children <18 years old in July 2018-June 2020. The 454 had a positive M. pneumoniae polymerase chain reaction assay. RESULTS: Most subjects had MRMP (78.4%), and all mutated strains had the A2063G transition. We defined MRMP* (n = 285) as MRMP pneumonia requiring admission and MSMP* (n = 72) as MSMP pneumonia requiring admission. Patients with MRMP pneumonia were older, more likely to have segmental/lobar pneumonia, and had more febrile days than those with MSMP pneumonia. C-reactive protein (CRP), lactate dehydrogenase (LDH), and percentage neutrophils were more strongly associated with MRMP* than MSMP* groups. Percentage neutrophils, CRP, and alanine aminotransferase significantly changed between admission and follow-up measurements in patients with MRMP* (P < 0.05). The duration of admission positively correlated with the number of febrile days after initiation of antibiotic medication and laboratory variables (white blood cell count, CRP, and aspartate aminotransferase [AST]) (P < 0.05). Random forest analysis indicated that the number of febrile days after initiation of antibiotic medication, AST, and percentage neutrophils at admission was over five. CONCLUSIONS: This study indicated that children with M. pneumoniae pneumonia with a higher number of febrile days after initiation of antibiotic medication, AST, and percentage neutrophils at admission were more likely to have prolonged admission duration.

Clinical role of M. pneumoniae typing antibody detected by chemiluminescent immunoassay in the diagnosis of Mycoplasma pneumoniae pneumonia in children.

OBJECTIVE: The levels of serum M. pneumoniae typing antibodies in children with community-acquired pneumonia (CAP) were detected by chemiluminescent immunoassay (CLIA) to explore the clinical role of M. pneumoniae typing antibody (MP-IgM, MP-IgG) in M. pneumoniae pneumonia. METHODS: A total of 387 Child patients with CAP diagnosed at the Pediatric outpatient department of Zengcheng Branch, Nanfang Hospital, Southern Medical University, were enrolled between January 2020 to December 2021 and divided into M. pneumoniae pneumonia (MPP) group (n = 210) and non-M. pneumoniae pneumonia (NMPP) group (n = 177). Firstly, Clinical data, full blood count (WBC, NEU%, LYM%, MONO%, EOS%, BASO%, RBC, HGB, PLT) and biochemical tests (AST, LDH, ɑ-HBDH, CK, CKMB, CRP, PCT, IL-6) as well as laboratory diagnostic tests (MP-IgM, MP-IgG) were compared between the two groups. Secondly, we assessed the correlation between the average level of M. pneumoniae typing antibody detected by CLIA and the titer of anti-M. pneumoniae antibody (MP-Ab) tested by passive agglutination (PA) method. Thirdly, receiver operating characteristic (ROC) curve for the MP-IgM and MP-IgG was examined to evaluate the value of diagnosing M. pneumoniae pneumonia. Finally, we follow-up 120 cases of MPP group and analysis medication results. RESULTS: (1) Mean age, runny nose, expectoration, LYM%, NEU%, HGB, AST, MP-IgM and MP-IgG were statistically significant in the MPP group and NMPP group (all P < 0.05). (2) Correlation analysis showed that MP-IgM average level was linearly associated with MP-Ab titer (R2 = 0.84) and MP-IgG average level was exponentially correlated with MP-Ab titer under 1:640 (R2 = 0.96). (3) The ROC curve of MP-IgM and MP-IgG were significantly different (both P < 0.001). A serum MP-IgM level above 1 S/CO and MP-IgG level above 14.15 AU/mL were significant predictors for M. pneumoniae pneumonia: area under the curve (AUC) of 0.810, 0.815; standard error (SE) of 0.021, 0.022; 95 % confidence interval (CI) of 0.768-0.852, 0.773-0.858; the diagnostic sensitivity of 74.3 %, 62.1 %; and specificity of 72.9 %, 87.0 %; respectively. (4) Of the 120 children with M. pneumoniae pneumonia followed up, 79 (65.8 %) cases took azithromycin and 68 (86.1 %) cases were recovered. CONCLUSIONS: A series of our studies shown that, CLIA, speedy and automated clinical examination method, has higher specificity and sensitivity for the quantitative detection of MP-IgM and MP-IgG, playing an important role of early diagnosis as well as prompt intervention to reduces macrolide-resistant strains and sequelae of children with M. pneumoniae pneumonia.

Pinocembrin Relieves Mycoplasma pneumoniae Infection­Induced Pneumonia in Mice Through the Inhibition of Oxidative Stress and Inflammatory Response.

Pneumonia is a serious infectious disease with increased morbidity and mortality worldwide. The M. pneumoniae is a major airway pathogen that mainly affects respiratory tract and ultimately leads to the development of pneumonia. The current exploration was aimed to uncover the beneficial properties of pinocembrin against the M. pneumoniae-triggered pneumonia in mice via its anti-inflammatory property. The pneumonia was stimulated to the BALB/c mice via infecting them with M. pneumoniae (100 µl) for 2 days through nasal drops and concomitantly treated with pinocembrin (10 mg/kg) for 3 days. The azithromycin (100 mg/kg) was used as a standard drug. Then the lung weight, nitric oxide, and myeloperoxidase (MPO) activity was assessed. The content of MDA, GSH, and SOD activity was scrutinized using kits. The total cells and DNA amount present in the bronchoalveolar lavage fluid (BALF) was assessed by standard methods. The IL-1, IL-6, IL-8, TNF-α, and TGF contents in the BALF samples and NF-κB level in the lung tissues were assessed using kits. The lung histopathology was assessed microscopically to detect the histological alterations. The 10 mg/kg of pinocembrin treatment substantially decreased the lung weight, nitric oxide (NO) level, and MPO activity. The MDA level was decreased, and GSH content and SOD activity were improved by the pinocembrin treatment. The pinocembrin administered pneumonia animals also demonstrated the decreased total cells, DNA amount, IL-1, IL-6, IL-8, TNF-α, and TGF in the BALF and NF-κB level. The findings of histological studies also witnessed the beneficial role of pinocembrin against M. pneumoniae-infected pneumonia. In conclusion, our findings confirmed that the pinocembrin effectively ameliorated the M. pneumoniae-provoked inflammation and oxidative stress in the pneumonia mice model. Hence, it could be a hopeful therapeutic agent to treat the pneumonia in the future.

Acute Pancreatitis: An Unusual Extrapulmonary Manifestation of Mycoplasma pneumoniae.

Mycoplasma pneumoniae is a respiratory pathogen responsible for community-acquired atypical pneumonia. Apart from respiratory manifestations, other system involvement has also been reported. We present a case of interstitial pneumonia and a concurrent episode of acute pancreatitis in a young female who presented with fever, cough, vomiting, and epigastric pain. The abdominal evaluation revealed epigastric tenderness with no signs of organomegaly. Her complete metabolic profile was nonsignificant except for elevated serum lipase and amylase. Clinical, serological, and radiological features and detailed investigations confirmed the diagnosis of acute pancreatitis and interstitial pneumonia caused by M. pneumoniae in the absence of any other etiology. Her respiratory and gastrointestinal symptoms improved rapidly after commencing clarithromycin, providing a possible link between M. pneumoniae and pancreatitis.

Geraniol relieves mycoplasma pneumonia infection-induced lung injury in mice through the regulation of ERK/JNK and NF-κB signaling pathways.

BACKGROUND: Pneumonia is a serious pediatric lung injury disease caused by Mycoplasma pneumoniae (M. pneumoniae) with increasing global prevalence every year. The WHO has reported that nearly 19% of children die due to pneumonia worldwide. OBJECTIVE: The present research was conducted to discover the ameliorative properties of geraniol against M. pneumoniae-provoked pneumonia in mice through the modulation of inflammatory responses. METHODOLOGY: The pneumonia was provoked in the male Swiss albino mice via infecting animals with 100 µl of M. pneumoniae for 2 days and supplemented concurrently with 20 mg/kg of geraniol for 3 days. 100 mg/kg of azithromycin was used as a standard drug. The nitric oxide (NO) level and MPO activity were measured using kits. The SOD activity, GSH, and MDA levels were studied using standard methods. The polymerase chain reaction (PCR) study was performed to examine the M. pneumoniae DNA load. The inflammatory cytokines status was assessed by assay kits. The ERK1/2, JNK1/2, and NF-κB expressions were studied by reverse-transcription (RT-PCR). The lung tissues were analyzed microscopically to investigate the histological alterations. RESULTS: Geraniol treatment effectively reduced lung weight, NO level, and MPO activity in the pneumonia mice. The total cells and M. pneumoniae DNA load were also decreased by the geraniol. The SOD activity and GSH level were improved and MDA was decreased by the geraniol treatment. The IL-1, IL-6, IL-8, TNF-α, and TGF status were appreciably depleted by the geraniol in the pneumonia mice. Geraniol also suppressed the ERK1/2 and NF-κB expressions in the lung tissues. Histological findings also suggest the therapeutic roles of geraniol against pneumonia in mice. CONCLUSION: In summary, our results proved the beneficial roles of geraniol against the M. pneumoniae-provoked pneumonia. Geraniol could be a hopeful therapeutic agent to treat pneumonia in the future.

ß-Caryophyllene ameliorates the Mycoplasmal pneumonia through the inhibition of NF-κB signal transduction in mice.

BACKGROUND: Pneumonia is a frequent infectious disease that mainly affects the children and the global death rate is nearly 19% among children at the below 5 age. ß-caryophyllene is an active compound, mainly occurs in the spices and it possesses immense biological activities. OBJECTIVE: This investigation deliberated to scrutinize the beneficial actions of ß-caryophyllene against the M. pneumoniae induced pneumonia. METHODS: The pneumonia was stimulated to the BALB/c mice by infecting them with 100 µl of M. pneumonia for 2 days via nasal drops with the concomitant treatment with 20 mg/kg of ß-caryophyllene. The total cells in the BALF of test mice were counted by using the Neuber chamber. The total protein and the pro-inflammatory cytokines status were examined by using the commercial ELISA kits. The PCR technique was used to measure the M. pneumoniae bacterial load. The NF-ƙB expression was investigated using western blotting. The lung tissues were analyzed microscopically. RESULTS: The ß-caryophyllene notably diminished the total protein status, total cell count, and bacterial load in the pneumonia provoked mice. The marked reduction in the status of pro-inflammatory regulators was seen in the ß-caryophyllene supplemented pneumonia mice. ß-caryophyllene also down-regulated the expression of NF-ƙB thereby reduced the lung inflammation and tissue damages as seen in the result of histological analysis. CONCLUSION: These findings were confirmed the therapeutic potential of ß-caryophyllene against the M. pneumoniae-activated pneumonia in animals.

Locating macromolecular assemblies in cells by 2D template matching with cisTEM.

For a more complete understanding of molecular mechanisms, it is important to study macromolecules and their assemblies in the broader context of the cell. This context can be visualized at nanometer resolution in three dimensions (3D) using electron cryo-tomography, which requires tilt series to be recorded and computationally aligned, currently limiting throughput. Additionally, the high-resolution signal preserved in the raw tomograms is currently limited by a number of technical difficulties, leading to an increased false-positive detection rate when using 3D template matching to find molecular complexes in tomograms. We have recently described a 2D template matching approach that addresses these issues by including high-resolution signal preserved in single-tilt images. A current limitation of this approach is the high computational cost that limits throughput. We describe here a GPU-accelerated implementation of 2D template matching in the image processing software cisTEM that allows for easy scaling and improves the accessibility of this approach. We apply 2D template matching to identify ribosomes in images of frozen-hydrated Mycoplasma pneumoniae cells with high precision and sensitivity, demonstrating that this is a versatile tool for in situ visual proteomics and in situ structure determination. We benchmark the results with 3D template matching of tomograms acquired on identical sample locations and identify strengths and weaknesses of both techniques, which offer complementary information about target localization and identity.

Analysis of mycoplasma pneumoniae infection among children with respiratory tract infections in hospital in Chengdu from 2014 to 2020.

BACKGROUND: Respiratory tract infection (RTIs) is one of common diseases among the children. In recent years, the incidence of mycoplasma pneumoniae (M. pneumoniae) infection rate has been increasing, which cause respiratory tract infection. This study sought to analyze the epidemiological characteristics of M. pneumoniae hospitalized children with RTIs to provide a theoretical basis for clinical diagnosis and treatments in Chengdu, China. METHODS: The data of 22,882 cases of children who had been hospitalized for RTIs were collected. M. pneumoniae immunoglobulin M (IgM) antibody was detected using the indirect immunofluorescence method and passive agglutination method. The demographic features of patients, clinical diagnoses and laboratory data were also analyzed. RESULTS: A total of 4,213 children tested positive for M. pneumoniae. The total positive rate was 18.41% (18.30% for males and 22.72% for females). Female children had statistically significant higher positive rates than male children (χ2=198.078, P<0.01). The positive rates of M. pneumoniae differed significantly among children of different ages (F=162.7532, P<0.01). The incidence rate of M. pneumoniae in 2017 and 2019 was significantly higher than the average (F=538.95, P<0.01). There were higher M. pneumoniae positive rates from April to May, and September to October (P<0.05) in 2016, 2017, 2018, and 2019. There was no correlation between M. pneumoniae infection and temperature and humidity (P>0.05). There was negative correlation with PM2.5 (particulate matter in the air <2.5 µm) (R=-0.293, P<0.01) and PM10 (particulate matter in the air <10 µm). (R=-0.285, P<0.01). There were significant differences in the constituent ratios of cases of M. pneumoniae infection between in 2020 and other years (F=159.35, P<0.01). Bronchopneumonia accounted for the highest proportion of cases, followed by acute bronchitis and the exacerbation of asthma in 2020. CONCLUSIONS: The epidemiological distribution of M. pneumoniae in children with RTIs in Chengdu was found to be related to gender, age, year and month; however, no relationship was found to temperature and humidity. There was a higher M. pneumoniae positive rate in children with bronchial pneumonia and asthma in cases. The prevention measures used to control Coronavirus Disease 2019 (COVID-19) also effectively controlled the M. pneumoniae infection rate.

Highly homogeneous microbial communities dominated by Mycoplasma pneumoniae instead of increased resistance to macrolide antibiotics is the characteristic of lower respiratory tract microbiome of children with refractory Mycoplasma pneumoniae pneumonia.

BACKGROUND: Although researchers have found that the microbiota changed during the lower respiratory tract (LRT) infection, little was known about the association between LRT microbiome and refractory M. pneumoniae pneumonia (RMPP). METHODS: From June 28th, 2019 to March 23rd, 2020, we enrolled fifty-two children diagnosed with RMPP or non-refractory M. pneumoniae pneumonia (NRMPP), and characterized the structure and function of microbiota in the bronchoalveolar lavage fluid (BALF) by metagenomic next generation sequencing (mNGS). RESULTS: Based on Bray-Curtis distance between samples, samples in RMPP group were highly homogeneous, and Shannon index in the RMPP group was much lower than NRMPP group while Simpson index, which presents the degree of dominance, was higher in RMPP group. The dominant taxon with relative abundance greater than 50% was merely Mycoplasma among RMPP and NRMPP patients, but the proportions of other taxonomic distribution were different. M. pneumoniae was the dominant species and occupied almost all niches in the vast majority of RMPP patients, whereas the other genera were dramatically lower. The NRMPP group was more enriched in antibiotic resistance genes (ARGs) than the RMPP group, and also exhibited a greater relative abundance of macrolide antibiotics resistance gene (macB) and fluoroquinolone antibiotic resistance genes (patA-B) in M. pneumoniae genome. In RMPP patients, higher relative abundance of Streptococcus pneumoniae had a strong correlation with increased hospitalization days while higher relative abundance of Streptococcus pneumoniae had a negative correlation with hospitalization days among NRMPP patients. CONCLUSIONS: The microbiota of LRT in children with RMPP was much more homogeneous and simpler than that of the NRMPP group and with lower relative abundance of macrolide antibiotics resistance gene in M. pneumoniae genome. M. pneumoniae was absolutely dominant in the vast majority of RMPP patients. Prolonged hospitalization days was associated with relative abundance of M. pneumoniae in NRMPP patients while it was related with other pathogens' relative abundance (e.g., Streptococcus pneumoniae) in RMPP patients.

Pseudomembranous laryngotracheobronchitis due to coinfection with human bocavirus 1 and Mycoplasma pneumoniae: a case report.

Pseudomembranous laryngotracheobronchitis is rarely reported yet potentially life-threatening infectious cause of airway obstruction in children. The causative organisms of this condition are often considered to promote bacterial superinfection following viral infection. We report a case of pseudomembranous laryngotracheobronchitis in a patient caused by human bocavirus 1 and Mycoplasma pneumoniae (M. pneumoniae). A 2-year-old child was admitted to our hospital presenting with cough, hoarseness, and labored breathing. Computed tomography of the chest revealed atelectasis of the right middle lobe of the lung with bronchostenosis and occlusion. Laryngeal edema, pseudomembrane formation and ulceration of the trachea were found during bronchoscopy. Chronic inflammation of the mucosa and local cellulose exudation with acute and chronic inflammatory cell infiltration were confirmed by hematoxylin-eosin staining. Human bocavirus 1 and M. pneumoniae were detected in the bronchoalveolar lavage fluid by next-generation sequencing. The patient tested positive for IgM antibodies against M. pneumoniae. Bronchoscopy was performed three times to clear the secretions in the airway, and azithromycin, ceftriaxone, methylprednisolone, budesonide inhalation, and ambroxol were administered as treatment. The patient's condition improved and she was discharged 21 days after admission. Clinicians should be aware of the potential involvement of human bocavirus 1 and M. pneumoniae in pseudomembranous laryngotracheobronchitis for accurate diagnosis and timely antibiotic administration, and to lower mortality and morbidity rates.

Evaluation of the CARDS toxin and its fragment for the serodiagnosis of Mycoplasma pneumoniae infections.

Mycoplasma pneumoniae (M. pneumoniae) is an important pathogen in community-acquired pneumonia. The community-acquired respiratory distress syndrome (CARDS) toxin is the only known virulence factor of M. pneumoniae. It is worth exploring whether this toxin can be used as a candidate antigen for the serodiagnosis of M. pneumoniae. In this study, the full-length, N-terminal, and C-terminal regions of the CARDS toxin were expressed and purified, and serological reactions were evaluated using ELISA. A total of 184 serum samples were collected and tested using a commercialized test kit. Eighty-seven samples were positive, and 97 samples were negative for infection. The purified recombinant proteins were used as antigens to test the serum via indirect ELISA. The sensitivity of the CARDS toxin, the N-terminal region, and the C-terminal region were 90.8%, 90.8%, and 92.0%, respectively. The specificity of the CARDS toxin, the N-terminal region, and the C-terminal region were 85.6%, 73.2%, and 93.8%, respectively. All three CARDS toxin proteins exhibited good reactivity, of which the C-terminal region had a good discrimination ability in human sera. This may have a potential diagnostic value for M. pneumoniae infections.

The effect of vibrating positive expiratory pressure therapy on refractory Mycoplasma pneumoniae pneumonia prognosis in children.

BACKGROUND: It is well documented that vibration techniques and positive expiratory pressure (PEP) can improve airway clearance, however, few clinical trials have studied the efficacy of vibrating PEP therapy on refractory Mycoplasma pneumoniae (M. pneumoniae) pneumonia (RMPP) in children. This study aimed to explore using vibrating PEP therapy in children with RMPP. METHODS: Sixty participants in the remission stage of RMPP were recruited into this randomized study and divided into two groups. The Acapella group (n=30) used the Acapella® Choice twice daily for 2 months to clear their airways. The control group (n=30) used traditional chest percussion or postural drainage to mediate sputum expectoration. The groups administered their respective treatments at home and were followed up weekly. Participants kept a record of their treatment and condition in a daily log. The primary outcome of this study was the resolution of chest images, and the secondary outcome was the sputum period. RESULTS: There was no significant difference in sex or age distribution between the two groups. There were no significant differences at baseline between the groups in the location of their lesions (P=0.11). After 2 months of treatment, there was a better resolution of chest images in the Acapella group (P=0.00) compared with the control group. Additionally, the mean sputum period was significantly shorter for the Acapella group than for the control group (Acapella: 7.97±1.54 days; control: 11.90±1.64 days; P=0.00). CONCLUSIONS: Vibrating PEP therapy is an effective therapy for children with RMPP, both in airway clearance and the resolution of lung abnormalities.

The mechanisms underlying antigenic variation and maintenance of genomic integrity in Mycoplasma pneumoniae and Mycoplasma genitalium.

Mycoplasma pneumoniae and Mycoplasma genitalium are important causative agents of infections in humans. Like all other mycoplasmas, these species possess genomes that are significantly smaller than that of other prokaryotes. Moreover, both organisms possess an exceptionally compact set of DNA recombination and repair-associated genes. These genes, however, are sufficient to generate antigenic variation by means of homologous recombination between specific repetitive genomic elements. At the same time, these mycoplasmas have likely evolved strategies to maintain the stability and integrity of their 'minimal' genomes. Previous studies have indicated that there are considerable differences between mycoplasmas and other bacteria in the composition of their DNA recombination and repair machinery. However, the complete repertoire of activities executed by the putative recombination and repair enzymes encoded by Mycoplasma species is not yet fully understood. In this paper, we review the current knowledge on the proteins that likely form part of the DNA repair and recombination pathways of two of the most clinically relevant Mycoplasma species, M. pneumoniae and M. genitalium. The characterization of these proteins will help to define the minimal enzymatic requirements for creating bacterial genetic diversity (antigenic variation) on the one hand, while maintaining genomic integrity on the other.

Epidemiology of macrolide-resistant Mycoplasma pneumoniae by age distribution in Japan.

INTRODUCTION: Mycoplasma pneumoniae (M. pneumoniae) is the major pathogen involved in community-acquired pneumonia in all age groups. Resistance to macrolides, the first-line treatment for M. pneumoniae infection, is a major global public health concern. However, studies evaluating macrolide-resistant M. pneumoniae infection simultaneously in all ages are limited. This study aimed to determine the prevalence and clinical characteristics of macrolide-resistant M. pneumoniae infection in terms of age distribution. METHODS: We enrolled 292 patients in Tokyo, Japan, who visited Eiju General Hospital or Zama Children's Clinic in 2015-2016. Patients were tested using real-time PCR for M. pneumoniae DNA. PCR-positive patients (n = 151) were further selected and sequentially divided into preschool-aged children (≤5 years, n = 31), school-aged children (6-15 years, n = 101), adolescents (16-19 years, n = 5), and adults (≥20 years, n = 14). We then analyzed the M. pneumoniae infection clinical characteristics, prevalence of macrolide-resistant infection, and 23S rRNA domain V resistance-associated mutation status. RESULTS: We found insignificant differences in the prevalence of macrolide-resistant M. pneumoniae infection among all groups, clinical characteristics, and resistance-associated mutation status in patients with macrolide-resistant M. pneumoniae infection. We also found statistically higher prevalence of mutation-positive (n = 85) M. pneumoniae in patients previously treated with macrolide compared to the mutation-negative group (n = 66); 63.8% vs 11.1% (p < 0.001), respectively. CONCLUSIONS: We found no significant differences in both clinical characteristics and prevalence of macrolide-resistant M. pneumoniae infection among all ages. Also, previous macrolide treatment contributes to drug-resistance.

Rational stepwise approach for Mycoplasma pneumoniae pneumonia in children.

Mycoplasma pneumoniae is a common pathogen that causes community-acquired pneumonia. In the past, M. pneumoniae was sensitive to macrolide antibiotics, and M. pneumoniae pneumonia (MPP) was usually a benign and self-limiting disease. However, despite use of the appropriate antibiotics, persistent fever and clinical deterioration may occur, leading to severe disease. Two major complicated conditions that may be clinically encountered are macrolide-resistant MPP and refractory MPP. Regarding the epidemics in Taiwan, before 2017, the mean rate of macrolide resistance was below 30%. Notably, since 2018, the prevalence of macrolide-resistant MPP in Taiwan has increased rapidly. Macrolide-resistant MPP shows persistent fever and/or no radiological regression to macrolide antibiotics and may even progress to severe and complicated pneumonia. Tetracyclines (doxycycline or minocycline) or fluoroquinolones are alternative treatments for macrolide-resistant MPP. Refractory MPP is characterized by an excessive immune response against the pathogen. In this context, corticosteroids have been suggested as an immunomodulator for downregulating the overactive host immune reaction. Overuse of macrolides may contribute to macrolide resistance, and thereafter, an increase in macrolide-resistant MPP. Delayed effective antimicrobial treatment is associated with prolonged and/or more severe disease. Thus, the appropriate prescription of antibiotics, as well as the rapid and accurate diagnosis of MPP, is important. The exact starting point, dose, and duration of the immunomodulator are yet to be established. We discuss these important issues in this review.

Mycoplasma Mucositis: A Case of Extrapulmonary Manifestation of Mycoplasma Pneumoniae.

We report the case of an 18-year-old male patient who presented for the evaluation of bilateral conjunctivitis, blurry vision of the left eye, penile lesions, and dysuria. The patient was admitted to the hospital due to widespread mucosal lesions and signs of disseminated infection. Laboratory studies revealed a leukocytosis of 17.41K/µL (normal: 4K/µL - 11K/µL) with a neutrophilic predominance of 82.7%. Chlamydia trachomatis, Neisseria gonorrhoeae, human immunodeficiency virus (HIV), antinuclear antibody (ANA), hepatitis, human leukocyte antigen B27 (HLA-B27), and pathergy test for Behcet's were all negative. Mycoplasma pneumoniae IgM and IgG, herpes simplex virus-1 (HSV-1) IgG and IgM, and HSV-2 IgG were all positive. It was determined that the cause for his lesions was likely Mycoplasma mucositis. He was treated with ceftriaxone, azithromycin, acyclovir, and methylprednisolone. After five days of treatment, complete resolution of symptoms was achieved and he was discharged home.