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

BACKGROUND: There is some evidence linking the infections with common organisms such as Chlamydophila pneumoniae, cytomegalovirus (CMV), Helicobacter pylori and HIV to myocardial infarction (MI). We had performed a serologic study to assess whether C.pneumoniae, CMV, H. pylori and HIV infections are associated with MI. METHODS: Serum samples were obtained from 54 cases of acute MI, 33 cases of old MI, and 60 normal controls. C-reactive protein (CRP) as an inflammation marker was measured and antibodies to C.pneumoniae, CMV, H.pylori and HIV were assayed by ELISA. Odds ratios (OR) were calculated against control group. RESULTS: CRP was significantly higher in the acute MI and old MI group. ORs of C.pneumoniae infection increased considerably in the acute MI (IgM 1.57, IgG 4.80) and old MI group (IgM 2.42, IgG 5.18). ORs of CMV infection were 3.30 in the acute MI and 5.12 in old MI group. ORs of H. pylori infection showed below 1 in the acute MI and old MI. Anti-HIV antibody showed all negative result in three groups, so OR could not be calculated. CONCLUSION: C.pneumoniae and CMV infections appear to be risk factors for MI.
Antibodies ; C-Reactive Protein ; Chlamydial Pneumonia* ; Chlamydophila pneumoniae* ; Chlamydophila* ; Coronary Artery Disease ; Cytomegalovirus* ; Enzyme-Linked Immunosorbent Assay ; Helicobacter pylori* ; Helicobacter* ; HIV Infections* ; HIV* ; Immunoglobulin G ; Inflammation ; Myocardial Infarction* ; Odds Ratio ; Risk Factors

OBJECTIVETo study the role of pathogenic microorganisms commonly associated with chronic eye disease, including Chlamydia psittaci, Chlamydia trachomatis, Chlamydia pneumoniae, herpes simplex virus (HSV) type 1 and type 2, and adenovirus type 8 and type 19, in the development of primary ocular adnexal mucosa-associated lymphoid tissue (MALT) lymphoma in Chinese patients.

METHODSSixty-eight archival cases of primary ocular adnexal lymphoproliferative lesions, including 38 cases of MALT lymphoma, 3 cases of non-MALT lymphoma and 27 cases of chronic inflammation, were enrolled into the study. DNA was extracted from the paraffin-embedded tissue samples. The presence of DNA of C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 were analyzed by multiplex touchdown enzyme time-release polymerase chain reaction (TETR-PCR).

RESULTSAll of the specimens yielded PCR products of over 100 base pairs and were thus suitable for TETR-PCR screening of infectious agents. The prevalence of DNA of C. psittaci, C. trachomatis and adenovirus type 19 were 0 in MALT lymphoma, non-MALT lymphoma and chronic inflammation. There were 2 cases positive for C. pneumoniae DNA, amongst the 38 cases of MALT lymphoma studied (5.3%, 2/38). HSV type 1, HSV type 2 and adenovirus type 8 DNA was found in each of the 3 patients with chronic inflammation.

CONCLUSIONThe study indicates that C. psittaci, C. trachomatis, C. pneumoniae, HSV type 1, HSV type 2, adenovirus type 8 and adenovirus type 19 probably play little role in the pathogenesis of ocular adnexal MALT lymphoma in Chinese patients.

Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; isolation & purification ; Chlamydia Infections ; microbiology ; Chlamydia trachomatis ; genetics ; isolation & purification ; Chlamydophila Infections ; microbiology ; Chlamydophila pneumoniae ; genetics ; isolation & purification ; Chlamydophila psittaci ; genetics ; isolation & purification ; DNA, Bacterial ; analysis ; DNA, Viral ; analysis ; Eye Infections ; microbiology ; virology ; Eye Neoplasms ; microbiology ; virology ; Herpes Simplex ; virology ; Herpesvirus 1, Human ; genetics ; isolation & purification ; Herpesvirus 2, Human ; genetics ; isolation & purification ; Humans ; Lymphoma, B-Cell, Marginal Zone ; microbiology ; virology ; Psittacosis ; microbiology

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

BACKGROUND: Chlamydia pneumoniae is an obligate intracellular organism, which can cause respiratory tract infections, exacerbation of asthma, and possibly atherosclerosis. Detection of C. pneumoniae in the atherosclerotic tissue was made using electron microscopy to elucidate the correlation between C. pneumoniae and atherosclerosis. METHODS: Ten cases of paraffin-embedded atherectomy tissue, which were positive in the immunohistochemistry (IHC) were prepared to demonstrate C. pneumoniae on the electron microscope. Two cases of atherosclerotic tissue, which were negative in IHC were processed identically at the same time as negative controls. The tissues were embedded in resin, ultrathin-sectioned, and stained with uranyl acetate and lead citrate, and then observed on electron microscopy. RESULTS: Nine of 10 (90%) atherosclerotic tissue samples were positive for C. pneumoniae on electron microscopy. Negative controls did not show elementary bodies. Elementary bodies of C. pneumoniae were rarely observed in a scattered pattern in the positive specimens. CONCLUSION: Elementary bodies of C. pneumoniae could be demonstrated on electron microscopy in 9 out of 10 atherosclerotic tissue samples with positive IHC for C. pneumoniae. This finding suggests that C. pneumoniae may play an important role in atherogenesis.
Asthma ; Atherectomy ; Atherosclerosis ; Chlamydia* ; Chlamydophila pneumoniae* ; Citric Acid ; Immunohistochemistry ; Microscopy, Electron* ; Pneumonia ; Respiratory Tract Infections

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

BACKGROUND: Chlamydia pneumoniae has recently been established as an important cause of acute respiratory tract infections such as pneumonia and bronchitis in humans. We introduced a 'touchdown' PCR method for detection of C. pneumoniae from sputum. METHODS: A total of 474 patients with respiratory infection were enrolled in the study. The sputum samples were tested for C. pneumoniae by the 'touchdown' PCR and cultured for Chlamydia. The sputum samples were pretreated with 5% NaOH for mucolysis. In 'touchdown' PCR, the first round PCR amplified DNA from both C. pneumoniae and Chlamydia psittaci, while the second round specifically targeted C. pneumoniae, allowing the two species to be differentiated. RESULTS: The 'touchdown' PCR could detect 10-2 inclusion forming unit (IFU) in the 1st round and 10-3 IFU in the second round PCR. None of the C. trachomatis serovars, C. psittaci and other organisms tested was amplified. 'Touchdown' PCR detected C. pneumoniae DNA in 24 (5%) of the 474 sputum samples. Nine patients with C. pneumoniae had community acquired pneumonia. Another nine patients had pulmonary tuberculosis of which three had coexisting pneumonia. Two patients had lung cancer, another two had chronic bronchitis, one had pharyngitis, and one person was a normal healthy individual. CONCLUSIONS: The sputum preparation with 5% NaOH and the 'touchdown' PCR method are effective in the detection of C. pneumoniae. C. pneumoniae is one of the most common causative agents for pulmonary infection.
Bronchitis ; Bronchitis, Chronic ; Chlamydia* ; Chlamydophila pneumoniae* ; Chlamydophila psittaci ; DNA ; Humans ; Lung Neoplasms ; Pharyngitis ; Pneumonia ; Polymerase Chain Reaction* ; Respiratory Tract Infections ; Sputum ; Tuberculosis, Pulmonary

Mycoplasma pneumoniae and Chlamydia pneumoniae have been suggested to take part in the acute exacerbation of bronchial asthma and chronic obstructive pulmonary disease (COPD). Several studies have questioned whether they may play pathogenic roles in connection with bronchial asthma and COPD. This study was designed to evaluate the seroprevalences of M. pneumoniae and C. pneumoniae in stable asthma and COPD patients, and to compare with control patients. The medical records of one hundred forty patients who underwent M. pneumoniae and C. pneumoniae serology were retrospectively reviewed. Seroprevalences of M. pneumoniae and C. pneumoniae in the asthma group (11.1% and 8.3%, respectively) were higher than in the control group (4.4% and 2.2%, respectively) without statistical significance. The seroprevalence of M. pneumoniae in the COPD group (16.9%) was significantly higher than in the control group, and the seroprevalence of C. pneumoniae in the COPD group (3.4%) was higher than in the control group without statistical significance. This study raises important questions about the relation of M. pneumoniae and C. pneumoniae infection with stable asthma or COPD.
Adult ; Aged ; Asthma/*microbiology ; Chlamydophila Infections/*epidemiology ; Chlamydophila pneumoniae/immunology ; Female ; Humans ; Male ; Middle Aged ; Pneumonia, Mycoplasma/*epidemiology ; Pulmonary Disease, Chronic Obstructive/*microbiology ; Seroepidemiologic Studies

OBJECTIVEThis paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.

METHODSMOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).

RESULTSIn Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II "healthy donors", 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.

CONCLUSIONThe novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.

Animals ; Antibodies, Monoclonal ; Bacterial Outer Membrane Proteins ; immunology ; Chlamydophila Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mice ; Protein Structure, Tertiary

OBJECTIVETo explore the relationship between Chlamydia pneumonia (CP) infection and hypertension as well as the interaction of traditional risk factors and CP infections on hypertension in Chinese Mongolian population.

METHODS1430 inhabitants living in Tongliao city, Inner Mongolia were selected as research subjects, including 488 hypertensives and 942 normotensives. Enzyme-linked immunoassay technique was used to test CP IgG antibody in the blood serum and SPSS 13.0 Microsoft was used to analyze the data.

RESULTSThe prevalence rates of hypertension were significantly different between CP IgG positive and negative groups, However, the trend was attenuated after adjusting age, sex, drinking, smoking, body mass index (BMI) and dyslipidemia. The CP infection rate was significant different in diastolic blood pressure groups, which was significantly higher in 80-84 mm Hg than that in the lowest DBP group. The interaction of chronic CP infection and other traditional risk factors were associated with hypertension in Chinese Mongolian ethnic groups. The ranking with significant sequence from high to low were BMI, dyslipidemia, alcohol intake, sex and age.

CONCLUSIONCP infection was not related to hypertension in Chinese Mongolian ethnic groups but the interaction with other traditional risk factors would increase the risk of developing hypertension.

China ; epidemiology ; Chlamydophila Infections ; epidemiology ; ethnology ; Chlamydophila pneumoniae ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hypertension ; epidemiology ; ethnology ; Immunoglobulin G ; analysis ; Male ; Mongolia ; ethnology ; Seroepidemiologic Studies