Objective To explore the clinical value of one-step visualization loop-mediated isothermal amplification(LAMP)in the detection of Mycoplasma pneumoniae(Mp). Methods One-step visualized LAMP,polymerase chain reaction(PCR),and enzyme-linked immunosorbent assay(ELISA)were used to simultaneously detect 108 clinical Mp specimens in children,which included 73 cases of Mp infection diagnosed by PCR and 35 cases of other chronic/acute respiratory tract infections.On the first day of admission,one-step visualization LAMP,PCR(fluorimetric method),and ELISA were used to test the throat swab and serum sample obtained from the same patient,and the Kappa value was calculated.The consistence between LAMP and PCR and that between LAMP and ELISA were compared.On the fifth day of admission,40 patients were resampled and the findings of these three tests on the first day and on the fifth day were compared. Results One-step visualization LAMP had a sensitivity of 100% and a specificity of 94.3%,whereas ELISA had a sensitivity of 65.8% and a specificity of 82.9%.The ratio of Kappa camparing one-step visualization LAMP and PCR was 0.956 and the ratio of Kappa camparing one-step visualization LAMP and ELISA was 0.38.The number of positive specimens detected by LAMP was higher than that by ELISA on the first day. Conclusions One-step visualization LAMP has excellent sensitivity and specificity in detecting early acute Mp infection.It has high consistency with PCR and can be applied to detect Mp.
Child ; Enzyme-Linked Immunosorbent Assay ; Humans ; Mycoplasma pneumoniae ; isolation & purification ; Nucleic Acid Amplification Techniques ; Pneumonia, Mycoplasma ; diagnosis ; Polymerase Chain Reaction ; Sensitivity and Specificity

OBJECTIVETo investigate the distribution characteristics of Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Legionella pneumophila (LP) in hospitalized children with acute respiratory tract infection (ARTI).

METHODSA total of 13 198 hospitalized children with ARTI were enrolled as study subjects. Whole blood and urine were collected. The passive agglutination was used to detect serum MP-IgM, ELISA was used to detect serum CP-IgM, and immunochromatography was performed to detect urinary LP antigen.

RESULTSAmong the 13 198 hospitalized ARTI children, the detection rates of MP, CP, and LP were 25.31%, 12.74% and 3.27%, suggesting that MP had the highest detection rate (P<0.0125). The detection rates of MP in 2013 and 2014 were significantly higher than that in 2012 (P<0.0125). CP had the highest detection rate in 2013, and LP had the highest detection rate in 2014 (P<0.0125). These three pathogens were detected all around the year, and MP had the highest detection rate in all seasons (P<0.0125). The detection rate of mixed infection with three pathogens was 4.35%, and mixed infection with MP and CP was the most common (P<0.0071). Among the children in different age groups, the patients aged 5-16 years showed the highest overall detection rate of three pathogens (P<0.0071). Among the children with different types of ARTI, the children with bronchopneumonia showed the highest overall detection rate of three pathogens (P<0.0045).

CONCLUSIONSMP, CP, and LP, particularly MP, are important pathogens for children with ARTI in the local area. LP infection tends to increase year by year and should be taken seriously in clinical practice.

Acute Disease ; Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; Chlamydophila pneumoniae ; isolation & purification ; Female ; Humans ; Infant ; Infant, Newborn ; Legionella pneumophila ; isolation & purification ; Mycoplasma pneumoniae ; isolation & purification ; Pregnancy ; Respiratory Tract Infections ; microbiology

No abstract available.
Acute Kidney Injury/microbiology ; Anti-Bacterial Agents/therapeutic use ; Biopsy ; Humans ; Kidney/*microbiology ; Male ; Middle Aged ; Mycoplasma pneumoniae/drug effects/*isolation & purification ; Nephritis/diagnosis/drug therapy/*microbiology ; Pneumonia, Mycoplasma/diagnosis/drug therapy/*microbiology ; Skin Diseases, Bacterial/diagnosis/drug therapy/*microbiology ; Steroids/therapeutic use ; Treatment Outcome ; Vasculitis/diagnosis/drug therapy/*microbiology

The role of atypical bacteria and the effect of antibiotic treatments in acute bronchitis are still not clear. This study was conducted at 22 hospitals (17 primary care clinics and 5 university hospitals) in Korea. Outpatients (aged > or = 18 yr) who had an acute illness with a new cough and sputum (< or = 30 days) were enrolled in 2013. Multiplex real-time polymerase chain reaction (RT-PCR) was used to detect five atypical bacteria. A total of 435 patients were diagnosed as having acute bronchitis (vs. probable pneumonia, n = 75), and 1.8% (n = 8) were positive for atypical pathogens (Bordetella pertussis, n = 3; B. parapertussis, n = 0; Mycoplasma pneumoniae, n = 1; Chlamydophila pneumoniae, n = 3; Legionella pneumophila, n = 1). Among clinical symptoms and signs, only post-tussive vomiting was more frequent in patients with atypical pathogens than those without (P = 0.024). In all, 72.2% of the enrolled patients received antibiotic treatment at their first visits, and beta-lactams (29.4%) and quinolones (20.5%) were the most commonly prescribed agents. In conclusion, our study demonstrates that the incidence of atypical pathogens is low in patients with acute bronchitis, and the rate of antibiotic prescriptions is high.
Anti-Bacterial Agents/therapeutic use ; Bordetella parapertussis/genetics/*isolation & purification ; Bordetella pertussis/genetics/*isolation & purification ; Bronchitis/drug therapy/*microbiology ; Chlamydophila pneumoniae/genetics/*isolation & purification ; Community-Acquired Infections/microbiology ; Female ; Humans ; Hypertension/complications ; Legionella pneumophila/genetics/*isolation & purification ; Male ; Middle Aged ; Mycoplasma pneumoniae/genetics/*isolation & purification ; Real-Time Polymerase Chain Reaction ; Republic of Korea ; Sputum/microbiology

OBJECTIVETo investigate the distribution of pathogens of children with community acquired pneumonia (CAP) from the Chongqing area.

METHODSNasopharyngeal specimens and blood specimens of 1 613 children with CAP were collected between January 2014 and December 2014 for bacterial culture and detection of 7 respiratory viruses and antibodies against Mycoplasma pneumoniae (MP).

RESULTSThe overall positive rate of bacteria was 50.22% (810 cases). Hemophilus parainfluenzae (40.8%), Streptococcus pneumonia (29.7%) and Moraxelle catarrhalis (7.3%) were the predominant ones. Among the viruses, the top detected virus was respiratory syncytial virus (RSV, 58.3%), followed by parainfluenza virus type3 (17.4%) and adenovirus (14.3%). A total of 481 cases (29.82%) were MP-positive. The co-infection rate was 32.18% (519 cases), and the mixed infections of bacteria and viruses were common (47.4%).

CONCLUSIONSRSV and Hemophilus parainfluenzae are the major pathogens of CAP in children from the Chongqing area. MP is also an important pathogen. The co-infection of bacteria and viruses is prevalent.

Adolescent ; Child ; Child, Preschool ; Community-Acquired Infections ; etiology ; Female ; Haemophilus parainfluenzae ; isolation & purification ; Hospitalization ; Humans ; Infant ; Male ; Mycoplasma pneumoniae ; isolation & purification ; Pneumonia ; etiology ; Respiratory Syncytial Viruses ; isolation & purification

Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Anti-Inflammatory Agents ; therapeutic use ; Azithromycin ; administration & dosage ; therapeutic use ; Biomarkers ; blood ; Child ; Eyelids ; pathology ; Humans ; Immunoglobulin M ; blood ; Lip ; pathology ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Mucositis ; diagnosis ; drug therapy ; microbiology ; Mycoplasma pneumoniae ; drug effects ; isolation & purification ; Pneumonia, Mycoplasma ; complications ; diagnosis ; drug therapy

OBJECTIVETo analyze the clinical characteristics of Mycoplasma pneumoniae-associated hemophagocytic syndrome (MP-HLH).

METHODA retrospective investigation of the clinical manifestation, laboratory test, imagelogy, clinical course and outcome of 3 cases with MP-HLH seen between June 2013 and July 2013 in Shenzhen Children's Hospital, and review of relevant literature were conducted.

RESULTOf the 3 cases of MP-HLH, 2 were males, one was female, the ages were 1 year, 3 years and 6 years, respectively. They had no underlying disease previously. All the 3 cases had onset of fever, cough as main symptoms. Diagnosis of refractory Mycoplasma pneumoniae pneumonia was made, which was accompanied by decreased neutrophils [(0.08-0.68)×10(9)/L], hemoglobin [(79-103) g/L], platelet [(64-157)×10(9)/L], plasma fibrinogen [(1.3-1.5) g/L], lactate dehydrogenase [(1,170-1,285) U/L] and increased serum ferritin [(936.7-39 789.0) µg/L] in the third week of course. In two cases the T lymphocytes decreased, and the NK cell activity decreased significantly in one. Bone marrow cytology showed prompted bone marrow hyperplasia, and the phenomenon of phagocytosed blood cells. CT scan was performed for all the cases and consolidation with pleural effusion were shown. Two cases were admitted to PICU, and required endotracheal intubation and mechanical ventilation. Flexible bronchoscopy and bronchial lavage were performed and bronchial cast was found in two cases. All of them were treated with macrolide combined with other antibiotics, glucocorticoids and gamma globulin combination therapy, including one case given dexamethasone [10 mg/(m2·d)], cyclosporine[6 mg/(kg·d)], etoposide [150 mg/(m2·d)] chemotherapy. Two cases were cured, and 1 case died. The authors summarized the 18 cases reported in domestic and foreign literature. Foreign children were diagnosed and treated with steroids in 1-2 weeks, and 10 cases were cured, and 2 cases died. They died of massive hemorrhage and meningoencephalitis, and domestic children were diagnosed and treated within two to 4 weeks after onset, 5 cases were cured, one case died of severe pneumonia.

CONCLUSIONMP-HLH is a rare disease in children, and had acute onset, rapid progression and high mortality. Early treatment with steroids was associated with a good prognosis, the key to successful treatment is early diagnosis and treatment, avoiding the immune cascade. Too late a diagnosis or development of serious complications may lead to death.

Anti-Bacterial Agents ; therapeutic use ; Bronchoalveolar Lavage Fluid ; Bronchoscopy ; Child ; Child, Preschool ; Fatal Outcome ; Female ; Fever ; Glucocorticoids ; therapeutic use ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; drug therapy ; microbiology ; Macrolides ; therapeutic use ; Male ; Mycoplasma pneumoniae ; isolation & purification ; Pleural Effusion ; Pneumonia, Mycoplasma ; complications ; diagnosis ; drug therapy ; Respiration, Artificial ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

We describe 2 cases of pneumonia caused by the same macrolide-resistant Mycoplasma pneumoniae in siblings. M. pneumoniae was identified using real-time PCR. Direct sequence analysis of the 23S rRNA gene revealed a point mutation in V domain (A2063G) of the 23S rRNA gene.
Anti-Bacterial Agents/pharmacology ; Child ; Child, Preschool ; Drug Resistance, Bacterial/drug effects ; Humans ; Macrolides/pharmacology ; Male ; Mutation ; Mycoplasma pneumoniae/*genetics/isolation & purification ; Pneumonia, Mycoplasma/*diagnosis/microbiology/radiography ; RNA, Ribosomal, 23S/*analysis ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, RNA ; Siblings

BACKGROUND: This study aimed to evaluate the prevalence of Mycoplasma pneumoniae in primary and tertiary care hospitals and its macrolide resistance rate. METHODS: Nasopharyngeal swabs were collected from 195 pediatric patients in primary and tertiary care hospitals from October to November 2010. The AccuPower MP real-time PCR kit (Bioneer, Korea) was used for the detection of M. pneumoniae. Direct amplicon sequencing was performed to detect point mutations conferring resistance to macrolides in the 23S rRNA gene. RESULTS: Among the 195 specimens, 17 (8.7%) were M. pneumoniae positive, and 3 of the strains (17.6%) obtained from these 17 specimens displayed the A2063G mutation in 23S rRNA. Three macrolide-resistant M. pneumoniae isolates were isolated from patients hospitalized at the primary care hospital. The positive rates of M. pneumoniae for the primary and tertiary care hospitals were 12.1% (15/124) and 2.8% (2/71), respectively (P=0.033). CONCLUSIONS: The positive rate of M. pneumoniae in the primary care hospital was higher than that in the tertiary care hospital. Simultaneous detection of M. pneumoniae and macrolide-resistant mutation genes in the 23S rRNA by real-time PCR is needed for rapid diagnosis and therapy of M. pneumoniae infections.
Anti-Bacterial Agents/*pharmacology ; Child, Preschool ; Drug Resistance, Bacterial/*drug effects ; Female ; Humans ; Infant ; Infant, Newborn ; Macrolides/*pharmacology ; Male ; Mycoplasma pneumoniae/genetics/*isolation & purification ; Nasopharynx/microbiology ; Pneumonia, Mycoplasma/epidemiology/microbiology ; Primary Health Care ; RNA, Ribosomal, 23S/analysis ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction ; Tertiary Healthcare

OBJECTIVETo elucidate the etiology of acute respiratory tract infection (ARI) in hospitalized children in Suzhou from 2005 to 2011.

METHODSA total of 10 243 hospitalized children with ARI in Children's Hospital Affiliated to Soochow University from September 2005 to October 2011 were enrolled in the study. The clinical information was collected; and the nasopharyngeal aspiration fluid and serum samples were sent for multi-pathogen detection. Respiratory syncytial virus (RSV), influenza virus type A and B (IV-A, IV-B), parainfluenza virus type 1-3 (PIV-1-PIV-3) and adenovirus (ADV) were detected by direct immunofluorescence assay. Human bocavirus (HBoV), mycoplasma pneumoniae (MP) and chlamydia pneumoniae (CP) were detected by fluorescent quantitative PCR while human metapneumovirus (hMPV) was detected by reverse transcription PCR (RT-PCR). Sputum culture was applied to detect bacterial infection and quantitative ELISA was adopted to detect the specific antibodies of MP and CP. The results of the above detections were analyzed, and thereby to explore the prevalent pathogens among different aging children and the seasonal distribution and characteristics of the disease.

RESULTSAt least one type of pathogen was detected in 5871 out of 10 243 hospitalized children and the overall positive rate was 57.32%; including 3326 virus samples with positive rate at 32.47% (3326/10 243), 2870 bacteria samples with positive rate at 28.02% (2870/10 243) and 2759 atypical pathogen samples,with positive rate at 26.94% (2759/10 243). MP was the most common pathogen,whose detected rate was 25.74% (2637/10 243). The median age of children with RSV (6 months) or PIV-3(8 months) infection was younger than the median age of all hospitalized children (12 months) (χ(2) = 380.992, 34.826, P < 0.05). While the median age of children with ADV (42 months), HBoV (14 months) or IV-A (24 months) infection was older than it of all hospitalized children (χ(2) = 83.583, 13.169, 18.012, P < 0.05). The median age of children with MP (30 months),streptococcus pneumoniae (17 months) or haemophilus parainfluenzae (21 months) infection was older than it of all hospitalized children (χ(2) = 728.299, 60.463, 8.803, P < 0.05). The detected rate of RSV in the groups of children aging less than 6 months, 7-12 months, 2-3 years, 4-5 years and over 6 years was separately 25.59% (840/3283), 17.05% (333/1953), 11.85% (310/2615), 6.68% (90/1347), and 2.87% (30/1045); which decreased while the age grew (χ(2) = 178.46, P < 0.01). Conversely, the positive rate of MP increased with the age growing (χ(2) = 379.21, P < 0.01). The rate in the above groups was 8.25% (271/3283), 19.46% (380/1953), 33.00% (863/2615), 41.43% (558/1347), 54.07% (565/1045), respectively. RSV and IV-A were prevalent in winter, whose detected rates were 35.73% (941/2634) and 4.44% (117/2634) respectively.hMPV infection was common in spring, with the detected rate at 10.55% (278/2634); while HBoV infection was common in summer and autumn, with the positive rate at 9.99% (149/1491) and 9.71% (98/1009). MP and CP were frequently detected in summer, up to 31.27% (819/2619) and 10.07% (43/427) respectively. RSV was the most common pathogen in bronchiolitis (33.27% (866/2603)) and MP was the most common pathogen in bronchopneumonia (26.05% (1152/4422)) and lober pneumonia (52.25% (267/511)).

CONCLUSIONMP and RSV were the most common pathogens in respiratory tract infection in hospitalized children. The novel virus included hMPV and HBoV, which also played an important role in ARI. Different pathogens were prevalent in different ages; with respective seasonal distribution and characteristics.

Acute Disease ; Adolescent ; Child ; Child, Hospitalized ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Mycoplasma pneumoniae ; isolation & purification ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; epidemiology ; microbiology ; virology ; Seasons