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

Mycoplasma infection is the most prevalent contamination in cell culture. Analysis of cell culture in laboratories from different countries shows that mycoplasma contamination ranges from 15% to 80% and, in some cases, even reaches 100% (Chernov et al., 2014). Whilst mycoplasma infection is not visible to the naked eye in cell culture, the consequences of mycoplasma contamination have been shown to induce a number of cellular changes, for example, increased resistance to chemotherapeutic drugs. Therefore, any results obtained from tissue culture studies, in the presence of mycoplasma contamination, potentially render the data invalid (Kim et al., 2015; Gedye et al., 2016). As such, mycoplasmas are not harmless bystanders and cannot be ignored in in vitro studies.
Arginine/pharmacology* ; HEK293 Cells ; Humans ; Mycoplasma/isolation & purification* ; Plasmids ; Transfection

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

Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control.
Animals ; Diagnostic Tests, Routine/methods/*veterinary ; Female ; Longitudinal Studies ; Mouth/microbiology ; Mycoplasma Infections/diagnosis/microbiology/*veterinary ; Mycoplasma hyorhinis/*isolation & purification ; Mycoplasma hyosynoviae/*isolation & purification ; Nose/microbiology ; Palatine Tonsil/microbiology ; Real-Time Polymerase Chain Reaction/*veterinary ; Reproducibility of Results ; Swine ; Swine Diseases/*diagnosis/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

This study examined the occurrence of Anaplasma spp. and hemoplasma infection in leopard cats, Prionailurus bengalensis euptilurus, in Korea. Twenty-nine biological samples were tested by molecular analysis. Two (6.9%) and eight (27.6%) tested specimens were positive for Anaplasma bovis and hemoplasma infection, respectively. Based on our results, Anaplasma/Ehrlichia spp. and hemoplasma are regularly infecting leopard cat populations of Korea. Considering their endangered status, regular monitoring of infection by arthropod-borne pathogens known to cause clinical symptoms in feline hosts such as Anaplasma/Ehrlichia spp. and hemoplasma would be crucial as part of ongoing conservation efforts.
Anaplasma/*isolation & purification ; Anaplasmosis/*epidemiology/microbiology ; Animals ; DNA, Bacterial/genetics ; *Felidae ; Molecular Sequence Data ; Mycoplasma/*isolation & purification ; Mycoplasma Infections/epidemiology/microbiology/*veterinary ; Phylogeny ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics ; Republic of Korea/epidemiology ; Sequence Analysis, DNA/veterinary

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

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

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

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chlamydia Infections/parasitology ; Chlamydia trachomatis/isolation & purification ; Coinfection ; Humans ; Male ; Middle Aged ; Mycoplasma Infections/parasitology ; Mycoplasma genitalium/isolation & purification ; *Polymerase Chain Reaction ; Prostatitis/epidemiology/*parasitology ; Republic of Korea/epidemiology ; Trichomonas Infections/*diagnosis ; Trichomonas vaginalis/*isolation & purification ; Young Adult