Chlamydophila psittaci/genetics* ; Humans ; Pneumonia ; Psittacosis/diagnosis*

The prevalence of feline herpesvirus-1 (FHV-1), feline calicivirus (FCV), and Chlamydophila (C.) felis was studied in cats of an animal shelter in Korea. Total 78 cats without ocular and upper respiratory tract disease were examined. Specimens were obtained from ocular conjunctiva and oropharynx. Using multiplex polymerase chain reaction (PCR) and reverse transcription PCR, three pathogens were simultaneously detected. In examined 78 cats, 49 (63%) cats were positive for FHV-1. However, all specimens were negative for C. felis and FCV. In conclusion, many cats recovered from FHV-1 infection remain subclinical carriers in shelter environment.
Animals ; Caliciviridae/genetics ; Caliciviridae Infections/epidemiology/*veterinary ; Cat Diseases/*epidemiology/*microbiology/*virology ; Cats ; Chlamydophila/genetics ; Chlamydophila Infections/epidemiology/*veterinary ; DNA Primers/genetics ; Herpesviridae/genetics ; Herpesviridae Infections/epidemiology/*veterinary ; Housing, Animal ; Korea/epidemiology ; Prevalence ; Reverse Transcriptase Polymerase Chain Reaction

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

The purified elementary bodies of C. pneumoniae TW-183 were used for immunization of male BALB/c mice, the spleen cells of these mice were fused with SP2/0 cells and the hybrid cells were cloned by limiting dilution. One clone that secreted the C. pneumoniae monoclonal antibody (Cpn-McAb) stably was obtained finally. The Cpn-McAb belonged to IgG2b class and anti-Cpn-MOMP; the outcome of micro-immunofluorescence showed its weak cross reaction with the C. psittaci elementary body but it has no cross reaction with C. trachoma elementary body. It has the same speciality of the imported Cpn-McAb. For the evaluation of Cpn-McAb, the peripheral blood mononuclear cell specimens of 454 patients were detected by self-made Cpn-McAb and imported Cpn-McAb at the same time. The positive rates of Cpn-antigen were 53.3% for self-made Cpn-McAb and 52.6% for imported Cpn-McAb,showing high concordance between them (Kappa=0.714). The results showed that self-made Cpn-McAb has almost the same high specificity and sensitivity as imported Cpn-McAb, so the self-made Cpn-McAb may replace imported Cpn-McAb to detect Cpn specific antigen and be helpful to diagnosing and treating the clinical diseases associated with Cpn infection.
Animals ; Antibodies, Bacterial ; immunology ; Antibodies, Monoclonal ; biosynthesis ; genetics ; Antibody Specificity ; Chlamydia Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; immunology ; Hybridomas ; secretion ; Male ; Mice ; Mice, Inbred BALB C

BACKGROUND: Differentiation of atypical pathogens is important for community-acquired pneumonia (CAP). In this study, we compared sputum and nasopharyngeal swabs (NPS) for use in detection of Mycoplasma pneumoniae (MP), Chlamydophila pneumoniae (CP), and Legionella pneumophila (LP), using Seeplex PneumoBacter ACE Detection Assay (PneumoBacter; Seegene). METHODS: Sputum and NPS specimens were collected from patients in 15 hospitals. DNA was extracted from sputum using QIAamp DNA Stool Mini Kit (Qiagen) and from NPS using easyMAG (bioMerieux). Both types of specimens were evaluated by multiplex PCR using PneumoBacter. To determine the diagnostic performance of this assay, sputum samples were also tested using BD ProbeTec ET Atypical Pneumonia Assay (APA; Becton Dickinson). RESULTS: Among 217 sputum and NPS, 20 (9.2%), 2 (0.9%), and 0 sputum were positive for MP, LP, and CP, respectively, whereas 8 (3.7%) NPS were positive for MP. The sputum APA test yielded 186, 206, and 204 interpretable results for MP, LP, and CP, respectively. Of these, 21 (11.3%) were positive for MP, 2 (1.0%) were positive for LP, and 0 samples were positive for CP. Compared to APA, the sensitivity and specificity of the sputum assay for MP were 95.2% and 100.0%, respectively, whereas for the NPS assay, these were 38.1% and 93.9%. Sputum testing was more sensitive than NPS testing (P=0.002). For LP and CP diagnosis, PneumoBacter and APA tests agreed 100%. CONCLUSIONS: Specimen type is crucial and sputum is preferred over NPS for simultaneous detection of MP, LP, and CP using multiplex PCR in CAP.
Chlamydophila Infections/diagnosis ; Chlamydophila pneumoniae/*genetics/isolation & purification ; Community-Acquired Infections/*diagnosis ; DNA, Bacterial/analysis/isolation & purification ; Humans ; Legionella pneumophila/*genetics/isolation & purification ; Legionnaires' Disease/diagnosis ; Multiplex Polymerase Chain Reaction ; Mycoplasma pneumoniae/*genetics/isolation & purification ; Nasopharynx/*microbiology ; Pneumonia, Mycoplasma/diagnosis ; Reagent Kits, Diagnostic ; Sputum/*microbiology

OBJECTIVEChlamydia psittaci is an avian respiratory pathogen and zoonotic agent. The wide prevalence of C. psittaci poses a threat to the poultry industry and its employees. However, few commercial kits are available for detecting avian antibodies excluding the in-house ELISA kit. In this study, we developed a novel ELISA kit for detecting antibodies against C. psittaci based on the N-terminal fragment of polymorphic outer membrane protein D (PmpD-N) as the coating antigen.

METHODSThe antigen concentrations, primary antibody, and cut-off value were determined and optimized. The ELISA, designated PmpD-N ELISA, was assessed for sensitivity, specificity, and concordance using sera samples from 48 experimentally infected and 168 uninfected SPF chickens.

RESULTSThe sensitivity and specificity of PmpD-N ELISA were 97.9%, 100%, respectively, while the concordance was 98.1% as compared to that of MOMP-ELISA. No cross-reaction with positive sera for other avian pathogens was found. Using PmpD-N ELISA, 799/836 clinical samples were positive, including 93.0% and 98.1% positivity in layers and broilers, respectively.

CONCLUSIONThese data indicate that indirect ELISA with PmpD-N as the antigen candidate is a promising approach for the surveillance of C. psittaci infection.

Animals ; Bacterial Proteins ; analysis ; Chickens ; Chlamydophila psittaci ; genetics ; immunology ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; veterinary ; Membrane Proteins ; analysis ; Poultry Diseases ; diagnosis ; microbiology ; Psittacosis ; diagnosis ; microbiology ; veterinary ; Sensitivity and Specificity

OBJECTIVETo study Chlamydia pneumoniae (C. pneumoniae) infection in 110 patients with respiratory tract infection admitted to our hospital from January to December 1995 in Nanjing.

METHODSSputum and throat swab specimens were taken and C. pneumoniae DNA was detected by using polymerase chain reaction (PCR) with the HM-1-HR-1 primer pair. At the same time, serum samples were taken and immunoglobulin G and M (IgG and IgM) fractions of antibodies to C. pneumoniae were studied by microimmunofluorescence test.

RESULTSPrevalence of specific IgG was 70% in patients with respiratory tract infection. Seventeen patients (15.5%) were serologically diagnosed as having recent C. pneumoniae infections and 12 patients (10.9%) had positive PCR in sputum and/or swab specimens. The total positive rate was 22.7% (25/110) detected by PCR combined with serological tests. Acute infection of C. pneumoniae was common in patients with asthma (57.1%), pneumonia (35.0%), COPD (25.9%) and bronchitis (25.0%). Clinical features between C. pneumoniae infection and non-C. pneumonia infection showed no significant differences.

CONCLUSIONSChlamydia pneumoniae is an important pathogen that causes infection of the human respiratory tract and attention should be drawn to this special illness.

Acute Disease ; Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Antibodies, Bacterial ; blood ; Chlamydophila pneumoniae ; genetics ; immunology ; DNA, Bacterial ; analysis ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Pneumonia, Bacterial ; blood ; microbiology ; Polymerase Chain Reaction

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

Local epidemiologic data on the etiologies of patients hospitalized with community-acquired pneumonia (CAP) is needed to develop guidelines for clinical practice. This study was conducted prospectively to determine the proportion of atypical bacterial pathogens in adults patients hospitalized with CAP in Korea between October 2001 and December 2002. Microbiological diagnosis was determined by serology for antibodies to Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneu-mophila. Nucleic acid of M. pneumoniae and C. pneumoniae in respiratory samples and Legionella antigen in urine samples were detected. The study population consisted of 126 patients (71 males, 55 females), averaging 54.6 yr (SD+/-17.8), whose paired sera were available. An etiologic diagnosis for atypical pathogens was made in 18 patients (14.3%): C. pneumoniae 9 (7.1%), M. pneumoniae 8 (6.3%), and L. pneumophila 3 patients (2.4%). Streptococcus preumoniae and other typical pathogens were isolated from 36 patients (28.6%). Of 126 patients, 16 (12.7%) were admitted to intensive care unit and atypical pathogens were identified in 5 patients (31.3%). Initial clinical features of patients with pneumonia due to atypical, typical or undetermined pathogens were indistinguishable. We conclude that atypical pathogens should be seriously considered in hospitalized patients with CAP, when initiating empiric treatment in Korea.
RNA, Ribosomal, 16S/genetics ; Prospective Studies ; Polymerase Chain Reaction ; Pneumonia, Bacterial/blood/*microbiology/urine ; Mycoplasma pneumoniae/genetics/immunology/*isolation & purification ; Middle Aged ; Male ; Legionella pneumophila/genetics/immunology/*isolation & purification ; Korea ; Humans ; Hospitalization/statistics & numerical data ; Female ; Community-Acquired Infections/microbiology ; Chlamydophila pneumoniae/genetics/immunology/*isolation & purification ; Antigens, Bacterial/urine ; Antibodies, Bacterial/blood ; Aged ; Adult

PURPOSE: This study was designed to investigate the change of peroxisome proliferator-activated receptor gamma (PPARgamma) after the infection of the human coronary artery smooth muscle cells (HCSMCs) with Chlamydia pneumoniae (C. pneumoniae) and the effect of PPARgamma agonist on the expression of PPARgamma of C. pneumoniae-infected HCSMCs. MATERIALS AND METHODS: To determine the effect of PPARgamma agonist on the proliferation of C. pneumoniae-infected HCSMCs, rosiglitazone at various concentrations was applied 1 hour before inoculation of HCSMCs. RESULTS: The expression of PPARgamma mRNA in HCSMCs increased from 3 hours after C. pneumoniae infection and reached that of noninfected HCSMCs at 24 hours (p < 0.05). The expression of PPARgamma protein in HCSMCs also increased from 3 hours after C. pneumoniae and persisted until 24 hours as compared with that of noninfected HCSMCs (p < 0.05). The pretreatment of HCSMCs with rosiglitazone followed by the infection with C. pneumoniae augmented the expression of PPARgamma mRNA and protein (p < 0.05) and decreased cell proliferation. CONCLUSION: Our results showed that the expression of PPARgamma increases in response to C. pneumoniae infection and rosiglitazone further augmented the expression of PPARgamma. It is suggested that rosiglitazone could ameliorate the chronic inflammation in the vessel wall induced by C. pneumoniae by augmenting PPARgamma expression.
Blotting, Western ; Cell Line ; Cell Proliferation/drug effects ; Chlamydophila pneumoniae/growth & development/*physiology ; Gene Expression Regulation/drug effects ; Humans ; Muscle, Smooth, Vascular/cytology/drug effects/metabolism ; Myocytes, Smooth Muscle/drug effects/*metabolism/microbiology ; PPAR gamma/genetics/*metabolism ; RNA, Messenger/genetics/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Thiazolidinediones/pharmacology