BACKGROUND/AIMS: To use serological and multiplex polymerase chain reaction (PCR) assays to examine sputum samples from patients experiencing acute exacerbation of chronic obstructive pulmonary disease (AECOPD) for the presence of atypical pathogens, including Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila. METHODS: From September 2012 to February 2014, 341 patients with AECOPD attending outpatient clinics were enrolled as part of a randomized, double-blind, multicenter study. A commercial enzyme-linked immunosorbent assay was used to measure serum immunoglobulin M (IgM) and IgG antibody titers on the first day of the study and at 36 days post-enrollment. Multiplex PCR was used to test sputum samples for the presence of atypical pathogens. A urinary antigen test for L. pneumophila was performed on the first day. RESULTS: Nineteen patients (5.6%) showed serological evidence of acute infection with M. pneumoniae. Also, one and seven patients (2%) showed serological evidence of acute infection with C. pneumoniae and L. pneumophila, respectively. All DNA samples were negative for M. pneumoniae, C. pneumoniae, and L. pneumophila according to PCR. Only one urine sample was positive for L. pneumophila antigen, but serologic evidence was lacking. CONCLUSIONS: Serological testing suggested that infection by atypical pathogens during AECOPD was relatively uncommon. In addition, PCR provided no direct evidence of infection by atypical pathogens. Thus, atypical pathogens may not be a major cause of AECOPD in South Korea.
Ambulatory Care Facilities ; Chlamydophila pneumoniae ; DNA ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Korea ; Legionella pneumophila ; Multiplex Polymerase Chain Reaction ; Mycoplasma pneumoniae ; Pneumonia ; Pneumonia, Mycoplasma ; Polymerase Chain Reaction* ; Pulmonary Disease, Chronic Obstructive* ; Serologic Tests ; Sputum

Respiratory infections, which are caused by airborne pathogens, are the most common disease of all ages worldwide. This study was conducted to characterize the airborne respiratory pathogens in the public facilities in Busan, South Korea. A total of 260 public facilities were investigated in 2017, 52 seasonal indoor air from 2 hospitals and 208 indoor air samples from 208 randomly selected daycare centers. Among respiratory pathogen, 8 viral pathogens including human adenovirus (HAdV), human bocavirus (HBoV), human rhinovirus (HRV), human parainfluenza virus (HPIV), human respiratory syncytial virus (HRSV), human metapneumovirus (HMPV), human coronavirus (HCoV) and influenza virus (IFV), and 3 bacterial pathogens including Mycoplasma pneumoniae, Bordetella pertussis, and Chlamydophila pneumoniae, were investigated by multiplex real-time reverse transcription polymerase chain reaction. Pathogens were detected in 9 cases (3.4%). Among 9 positive samples, 6 (2.3%) cases were positive for HBoV and 3 (1.2%) cases were positive for IFV. All the positive cases were detected in daycare centers. Additionally, the concentration of HBoV was determined. In HBoV-positive samples, the cycle threshold (Ct) values of HBoV were 29.73~36.84, which are corresponding to the viral concentration of 4.91 × 10⁰ ~ 9.57 × 10² copies/ml. Serotype distribution of isolated HBoV was analyzed by sequencing of VP1/VP2 gene. All of the HBoV isolates were identified as HBoV type 1 with a high similarity among the isolates (>97%). No bacterial pathogen was identified in indoor air samples. Although virus concentration was not high in public facilities (daycare center), the presence of respiratory viral pathogens has been identified. Effective ventilation and air purification strategies are needed to reduce the indoor concentration of respiratory pathogens. A long-term and ongoing surveillance plan for respiratory pathogen management should be established.
Adenoviruses, Human ; Bordetella pertussis ; Busan ; Chlamydial Pneumonia ; Chlamydophila pneumoniae ; Coronavirus ; Human bocavirus ; Humans ; Korea ; Metapneumovirus ; Mycoplasma pneumoniae ; Orthomyxoviridae ; Paramyxoviridae Infections ; Pneumonia, Mycoplasma ; Polymerase Chain Reaction ; Public Facilities* ; Respiratory Syncytial Virus, Human ; Respiratory Tract Infections ; Reverse Transcription ; Rhinovirus ; Seasons ; Serogroup ; Ventilation

PURPOSE: Epitope spreading is a phenomenon in which distinct subdominant epitopes become major targets of the immune response. Heat shock protein (HSP) 60 from Porphyromonas gingivalis (PgHSP60) and peptide 19 from PgHSP60 (Pep19) are immunodominant epitopes in autoimmune disease patients, including those with periodontitis. It remains unclear whether Pep19 is a dominant epitope in subjects without periodontitis or autoimmune disease. The purpose of this study was to determine the epitope spreading pattern and verify Pep19 as an immunodominant epitope in healthy teenagers using dot immunoblot analysis. The patterns of epitope spreading in age-matched patients with type 1 diabetes mellitus (type 1 DM) and healthy 20- to 29-year old subjects were compared with those of healthy teenagers. METHODS: Peptide from PgHSP60, Mycobacterium tuberculosis HSP60 (MtHSP60), and Chlamydia pneumoniae HSP60 (CpHSP60) was synthesized for comparative recognition by sera from healthy subjects and patients with autoimmune disease (type 1 DM). Dot immunoblot analysis against a panel of peptides of PgHSP60 and human HSP60 (HuHSP60) was performed to identify epitope spreading, and a densitometric image analysis was conducted. RESULTS: Of the peptide from PgHSP60, MtHSP60, and CpHSP60, PgHSP60 was the predominant epitope and was most consistently recognized by the serum samples of healthy teenagers. Most sera from healthy subjects and patients with type 1 DM reacted more strongly with PgHSP60 and Pep19 than the other peptides. The relative intensity of antibody reactivity to Pep19 was higher in the type 1 DM group than in the healthy groups. CONCLUSIONS: Pep19 is an immunodominant epitope, not only in autoimmune disease patients, but also in healthy young subjects, as evidenced by their robust immunoreactivity. This result suggests that the Pep19-specific immune response may be an initiator that triggers autoimmune diseases.
Adolescent* ; Autoimmune Diseases ; Autoimmunity ; Chlamydophila pneumoniae ; Diabetes Mellitus, Type 1 ; Epitopes ; Healthy Volunteers ; Heat-Shock Proteins ; Humans ; Immunodominant Epitopes ; Mycobacterium tuberculosis ; Peptides ; Periodontitis ; Porphyromonas gingivalis* ; Porphyromonas*

BACKGROUND: Infection by the intracellular bacteria Mycoplasma pneumoniae, Chamydophila pneumoniae, and Legionella pneumophila are common causes of community-acquired pneumonia (CAP). This study describes the evaluation of a new multiplex real-time PCR test, EuDx™-PN MLC Detection Kit (EUDIPIA), which allows the simultaneous detection of M. pneumoniae, C. pneumoniae, and L. pneumophila in respiratory samples. METHODS: A total of 353 samples were tested using three PCR kits: multiplex PCR (Seeplex PneumoBacter ACE Detection Kit) and two multiplex real-time PCR (EuDx™-PN MLC Detection Kit and Anyplex™ II RB5 Detection Kit). The results were considered true positives (expanded standard) for M. pneumoniae, C. pneumoniae, and L. pneumophila if they were positive according to any of the three tests. RESULTS: The sensitivity and specificity of EuDx™-PN MLC Detection Kit were 93.3–100% and 100%, respectively. The agreement rate and Cohen's kappa coefficient (value) between EuDx™-PN MLC Detection Kit and Anyplex™ II RB5 Detection Kit for M. pneumoniae, C. pneumoniae, and L. pneumophila were 70–100% and 0.82–1, respectively. CONCLUSION: These results demonstrate that the EuDx™-PN MLC Detection Kit is a sensitive, specific, and useful screening tool for the detection of atypical pathogens in respiratory samples and can be helpful in selecting appropriate antimicrobial therapy for patients with respiratory infection.
Bacteria ; Chlamydial Pneumonia* ; Chlamydophila pneumoniae* ; Chlamydophila* ; Humans ; Legionella pneumophila* ; Legionella* ; Mass Screening ; Multiplex Polymerase Chain Reaction ; Mycoplasma pneumoniae* ; Mycoplasma* ; Pneumonia ; Pneumonia, Mycoplasma* ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Respiratory Tract Infections ; 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

PURPOSE: Chlamydia pneumoniae is a common intracellular bacterial pathogen and plays an important role in acute respiratory infections. The purpose of this study was to investigate clinical presentations of C. pneumoniae in children with acute respiratory infections. METHODS: We examined the medical records of pediatric patients (age<18 years) admitted with acute respiratory infections of C. pneumoniae to Gachon University Gil Medical Center between March 1, 2011 and August 31, 2014. We compared the clinical features of C. pneumoniae infection with that of Mycoplasma pneumoniae infection. RESULTS: We confirmed acute respiratory infections of C. pneumoniae in 110 patients out of 2,156 patients (5.1%) admitted with acute respiratory infections. The mean age was 37.2+/-30.1 months. More than half of them (54.5%) had coinfection. C. pneumoniae infection had mild and subacute courses. The mean duration of symptoms prior to admission was 8.5+/-13.8 days. There were remarkable seasonal variations and prevalence was higher in December and April (P=0.03 and P=0.02, respectively). Although rhinorrhea and pharyngeal injection were more common in C. pneumoniae infection (P<0.05), clinical signs and symptoms were similar between C. pneumoniae and M. pneumoniae. Extrapulmonary manifestations such as skin lesion, Gastrointestinal symptoms, hepatitis, and neurologic symptoms were common (41.0%) in C. pneumoniae infection and, had similar incidence in M. pneumoniae infection. CONCLUSION: C. pneumoniae is an important infectious agent of acute respiratory infections in children. Clinical pictures of C. pneumoniae are similar to M. pneumoniae, even in extrapulmonary manifestations. C. pneumoniae should be taken into consideration in differential diagnosis of acute respiratory infection in children.
Child* ; Chlamydia* ; Chlamydophila pneumoniae* ; Coinfection ; Diagnosis, Differential ; Hepatitis ; Humans ; Incidence ; Medical Records ; Mycoplasma pneumoniae ; Mycoplasma* ; Neurologic Manifestations ; Pneumonia ; Pneumonia, Mycoplasma* ; Prevalence ; Respiratory Tract Infections* ; Seasons ; Skin

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

Paroxysmal cold hemoglobinuria (PCH) is a rare diagnosis of acquired hemolytic anemia in children, which is caused by a specific cold antibody named Donath-Landsteiner hemolysin. Although various bacteria or viruses were reported as triggering factor of PCH, childhood PCH related to Chlamydia pneumoniae infection is uncommon. The authors report a case of childhood PCH which is related with suspicious Chlamydia pneumoniae infection, with a review of pertinent literature.
Anemia, Hemolytic ; Bacteria ; Child ; Chlamydia Infections ; Chlamydophila pneumoniae ; Diagnosis ; Hemoglobinuria, Paroxysmal ; Humans

OBJECTIVESTo study the correlation between Chlamydiae pneumonia and carotid atherosclerosis, and the correlation between the infection of Chlamydiae pneumonia and ischemic events.

METHODSThe study group consisted of 19 patients who underwent unilateral carotid endarterectomy surgery during the period from January 2010 to December 2011, and the atherosclerotic plaque specimens were harvested from these patients. The control group consisted of 10 patients who underwent extracranial-intracranial bypass surgery during the same period, and the normal external carotid artery specimens were got from these patients. The clinical data between the two groups had comparability. The presence of Chlamydiae pneumonia in atherosclerotic plaque and normal artery tissue were investigated by immunohistochemistry. The expression of Toll-like receptor 2 (TLR2), tumor necrosis factor-α (TNF-α) and vascular cell adhesion molecule-1 (VCAM-1) in the atherosclerotic plaque infected with Chlamydiae pneumonia were also detected. Data were analyzed using Fisher's exact test.

RESULTChlamydiae pneumonia was found in 9 of 19 atherosclerotic plaques, while no positive result was found in control group. The statistical analysis showed a significant difference (P = 0.011). Among the 19 patients in study group, 15 of them had ischemic events, and Chlamydiae pneumonia was found in 9 of these 15 patients; while the other 4 patients didn't have any ischemic events and no Chlamydiae pneumonia was found in them, but there was no statistical different between them (P = 0.087). Through immunohistochemistry, the expression of Chlamydiae pneumonia, TLR2, TNF-α and VCAM-1 were found in same area.

CONCLUSIONSThere is a correlation between Chlamydiae pneumonia and carotid atherosclerosis.And there might be a correlation between Chlamydiae pneumonia and cerebral ischemic events.

Carotid Artery Diseases ; metabolism ; microbiology ; Case-Control Studies ; Chlamydophila Infections ; complications ; Chlamydophila pneumoniae ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Plaque, Atherosclerotic ; microbiology ; Toll-Like Receptor 2 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

BACKGROUND AND OBJECTIVES: Chlamydia pneumoniae (C. pneumoniae) is a well-known pathogen of upper and lower respiratory tract infection. For a more efficient and practical cell culture system, we studied the growth of two clinical isolates of C. pneumoniae in selected cell lines derived from the human respiratory tract. MATERIALS AND METHOD: HeLa 229, HEp-2, which are well-known cell lines for the culture of C. pneumoniae, and AMC-HN-4, AMC-HN-7, AMC-HN-8, which are the newly developed cell lines in Korea were examined. Strains of C. pneumoniae used in this study were TW-183 and LKK-1 (the first Korean strain). Chlamydia was inoculated on each confluent cell line and incubated for 48 hrs. After staining with anti-Chlamydial lipopolysaccharide monoclonal antibody, we compared the efficiency of the C. pneumoniae infection on each cell line by counting the inclusion bodies. RESULTS: In culturing C. pneumoniae LKK-1, AMC-HN-4 cells consistently yielded higher inclusion body counts than HeLa 229 cells did, whereas inclusion body counts by AMC-HN-7 cells was low. AMC-HN-7, AMC HN-8 cells yielded lower inclusion body counts than HEp-2 cells. In culturing C. pneumoniae TW-183, AMC-HN-4, AMC-HN-7, and AMC-HN-8 cells did not yield lower inclusion body counts than HeLa 229 cells did. AMC-HN-7 cells yielded lower inclusion body counts than HEp-2 cells. CONCLUSION: The newly established upper airway epithelial cell lines, AMC HN-4 and AMC HN-8, had similar culture efficiency as HeLa 229 and HEp-2 cells for Chlamydial infection; therefore, these two cell lines could be used for the future studies of C. pneumoniae.
Cell Culture Techniques ; Cell Line ; Chlamydia ; Chlamydophila pneumoniae ; Epithelial Cells ; Epithelium ; Humans ; Inclusion Bodies ; Korea ; Pneumonia ; Respiratory Tract Infections