OBJECTIVETo investigate the expression of IFN-λ1 in respiratory epithelial cells of children with respiratory syncytial virus (RSV) infection and its relationship with RSV load.

METHODSThe nasopharyngeal swabs were collected from the children who were hospitalized with respiratory tract infection from June 2015 to June 2016. A direct immunofluorescence assay was used to detect the antigens of seven common respiratory viruses (including RSV) in the nasopharyngeal swabs. A total of 120 children who were only RSV positive were selected as the RSV infection group. A total of 50 children who had negative results in the detection of all viral antigens were selected as the healthy control group. Fluorescence quantitative real-time PCR was used to determine the RSV load and the expression of IFN-λ1 mRNA in the nasopharyngeal swabs of children in the two groups.

RESULTSThe expression of IFN-λ1 in the RSV infection group was significantly higher than that in the healthy control group (P<0.05). The expression of IFN-λ1 was positively correlated with RSV load (r=0.56, P<0.05).

CONCLUSIONSRSV can induce the expression of IFN-λ1 in respiratory epithelial cells, suggesting that IFN-λ1 may play an important role in anti-RSV infection.

Antigens, Viral ; analysis ; Child, Preschool ; Epithelial Cells ; immunology ; Female ; Humans ; Infant ; Infant, Newborn ; Interleukins ; analysis ; physiology ; Male ; Nasopharynx ; microbiology ; Real-Time Polymerase Chain Reaction ; Respiratory Syncytial Virus Infections ; immunology ; virology ; Viral Load

BACKGROUNDThe accuracy of nasopharyngeal aspirate (NPA) specimens in detecting lower respiratory pathogens remains controversial. The objective of this study was to evaluate the diagnostic accuracy of aspirates (NPAs) specimen in lower respiratory tract infections (LRTIs) in children.

METHODSThe prospective study was designed to collect the data of paired NPAs and bronchoalveolar lavage fluids from children with acute LRTIs from January 2013 to December 2015. All specimens were subjected to pathogen detection: bacterial detection by culture, Mycoplasma pneumoniae (Mp) detection by polymerase chain reaction assay and virus (influenza A and B viruses, parainfluenza virus [PIV] Types 1 and 3, respiratory syncytial virus, and adenovirus) detection by immunofluorescence assay. The diagnostic accuracy analysis of NPAs was stratified by age ≤3 years (n = 194) and >3 years (n = 294).

RESULTSWe collected paired specimens from 488 children. The positive rate of pathogen was 61.6%. For Streptococcus pneumoniae, NPA culture had the specificity of 89.9% and negative predictive value of 100% in age ≤3 years, the specificity of 97.2% and negative predictive value of 98.9% in age >3 years. For Mp, the positive predictive values of NPA was 77.4% in children ≤3 years, and 89.1% in children >3 years. For PIV III, NPA specimen had the specificity of 99.8% and negative predictive value of 96.5% in children ≤3 years. For adenovirus, NPA had the specificity of 97.8% and negative predictive value of 98.4% in age ≤3 years, the specificity of 98.9% and negative predictive value of 99.3% in age >3 years.

CONCLUSIONSNPAs are less invasive diagnostic respiratory specimens, a negative NPA result is helpful in "rule out" lower airway infection; however, a positive result does not reliably "rule in" the presence of pathogens.

Acinetobacter baumannii ; isolation & purification ; pathogenicity ; Adolescent ; Child ; Child, Preschool ; Clinical Laboratory Techniques ; methods ; Enterobacter aerogenes ; isolation & purification ; pathogenicity ; Escherichia coli ; isolation & purification ; pathogenicity ; Female ; Haemophilus influenzae ; isolation & purification ; pathogenicity ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Prospective Studies ; Pseudomonas aeruginosa ; isolation & purification ; pathogenicity ; Respiratory Tract Infections ; diagnosis ; microbiology ; Sensitivity and Specificity ; Staphylococcus aureus ; isolation & purification ; pathogenicity ; Streptococcus pneumoniae ; isolation & purification ; pathogenicity

OBJECTIVETo investigate the prevalence of Moraxella catarrhalis in the nasopharyngeal region of children with respiratory infection and the sensitivity of Moraxella catarrhalis isolates to common antimicrobial drugs.

METHODSNasopharyngeal swabs were collected from 1 082 children with respiratory infection, and Moraxella catarrhalis strains were isolated. The E-test method and disc diffusion test were used to determine the sensitivity of these strains to 11 common antimicrobial drugs. The test results were interpreted with reference to the standards of European Committee on Antimicrobial Susceptibility Testing (EUCAST), Clinical and Laboratory Standards Institute (CLSI), and British Society for Antimicrobial Chemotherapy (BSAC). The nitrocefin disc method was used to detect whether the isolated strains produced β-lactamase.

RESULTSAmong the 1 082 children with respiratory infection, 77 (77/1 082, 7.12%) carried Moraxella catarrhalis in the nasopharyngeal region. All the strains produced β-lactamase. With reference to all the three standards, all the strains were sensitive to amoxycillin-clavulanate and had a susceptibility rate of >95% towards ciprofloxacin and tetracycline. According to the EUCAST and CLSI standards, the susceptibility rate of the strains towards sulfamethoxazole-trimethoprim was as high as 98.7%, and more than 80% of all strains were sensitive to the three cephalosporins detected; however, with reference to the BSAC standard, only 2.6% of the strains were sensitive to cefuroxime, with an intermediate rate of 44.2% and a drug resistance rate of 53.2%. The rate of resistance to ampicillin was 81.8%. According to the CLSI standard, the non-susceptibility rate of the strains to erythromycin was 79.2%, and according to the EUCAST or BSAC standards, their non-susceptibility rate reached 90.9%; more than one third of the strains (27/77, 35.1%) had a minimal inhibitory concentration of >256 mg/L.

CONCLUSIONSAll of the Moraxella catarrhalis isolates in the nasopharyngeal region of children with respiratory infection produce β-lactamase and are sensitive to amoxycillin-clavulanate. These isolates have high susceptibility rates to the third- and fourth-generation cephalosporins and sulfamethoxazole-trimethoprim, but most of the isolates are resistant to ampicillin, cefuroxime, and erythromycin.

Adolescent ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Moraxella (Branhamella) catarrhalis ; drug effects ; isolation & purification ; Nasopharynx ; microbiology ; Respiratory Tract Infections ; microbiology

OBJECTIVETo compare the detection rates of Mycoplasma pneumoniae (MP) from nasopharyngeal aspirates (NPA) and bronchoalveolar lavage fluid (BALF) in children with pneumonia.

METHODSA total of 164 hospitalized children with pneumonia were enrolled. NPA and BALF of these children were collected within 24 hours of admission, and MP-DNA was detected by fluorescence quantitative PCR. Venous blood samples of all these children were collected within 24 hours of admission and on days 7-10 of treatment, and serum MP-IgM was detected using ELISA.

RESULTSThe positive rate of MP-DNA in NAP of the 164 cases was 51.8% , which was lower than 63.4% as the detection rate of MP-IgM in serum (P=0.044), and the two detection rates were moderately consistent with each other (Kappa=0.618, P<0.01). The positive rate of MP in BALF was 71.3%, which was not significantly different with that of MP-IgM in serum (P>0.05), and the detection rates were well consistent (Kappa=0.793, P<0.01). The detection rate of MP in NPA was lower than that in BALF (P<0.01), with moderate consistency between two of them (Kappa=0.529, P<0.01). The median MP copy number in BALF was significantly higher than that in NPA (P<0.01). The MP detection rates in NPA and BALF were significantly different among different courses of disease (P<0.05). As the course of disease extended, the MP detection rates in both NPA and BALF showed a declining trend; children with MP pneumonia of 1-2 weeks' duration and 2-4 weeks' duration had a higher MP-DNA detection rate in BALF than in NPA (P<0.05).

CONCLUSIONSMP-DNA in BALF has a high sensitivity, with a great significance for early diagnosis of MP pneumonia, while NPA MP-DNA tests may lead to a missed diagnosis.

Adolescent ; Bronchoalveolar Lavage Fluid ; microbiology ; Child ; Child, Preschool ; DNA, Bacterial ; analysis ; Female ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Pneumonia, Mycoplasma ; diagnosis

OBJECTIVETo investigate nasopharyngeal carriage rate, antimicrobial resistance and serotype distribution of Streptococcus pneumoniae among children with upper respiratory infection.

METHODSNasopharygeal swabs were collected from children with upper respiratory infection visiting the outpatient department of Beijing Children′s Hospital between March 2013 and February 2014. The antibiotic susceptibility was tested by Etest method, and the serotype was determined by Quellung reaction.

RESULTSThe nasopharyngeal carriage rate for Streptococcus pneumoniae was 23.8% (699/2 941). One hundred isolates were randomly chosen for antimicrobial susceptiblity test and serotyping. Up to 98.0% isolates were susceptible to parenteral penicillin. The susceptible rate against oral penicillin, however, was 33.0%. The non-susceptible rate to erythromycin and azithromycin was 97.0%. The multi-drug resistance rate was up to 86.0%. The common serotypes were 6A(12.0%), 19F(12.0%), 6B(10.0%), 23F(9.0%) and 14(8.0%). The coverage rates of 7-, 10- and 13-valent pneumococcal conjugate vaccine were 41.0%, 42.0% and 59.0% respectively.

CONCLUSIONSAbout 25% of children with upper respiratory infection are nasopharyngeal colonized by Streptococcus pneumoniae. The isolates show a high antimicrobial resistance. The 13-valent pneumococcal conjugate vaccine covers about 60.0% of the isolates.

Adolescent ; Carrier State ; epidemiology ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Humans ; Infant ; Male ; Nasopharynx ; microbiology ; Pneumococcal Vaccines ; immunology ; Respiratory Tract Infections ; microbiology ; Serotyping ; Streptococcus pneumoniae ; classification ; drug effects ; isolation & purification

OBJECTIVE: To research the prevalences of four kinds of bacteria including Alloiococcus otitidis, Streptococcus pneumonia, Haemophilus influenzae, and Moraxella catarrhalis in children with chronic otitis media with effusion (SOM) of the middle ear effusion, and the reproduction of the nasopharynx, so as to explore their meaning for the children with SOM. METHOD: Alloiococcus otitidis, Streptococcus pneumonia, Haemophilus influenza, and Moraxella catarrhal were investigated in the samples obtained from middle ear effusion and nasopharyn- geal swabs, using PCR and conventional bacterial culture methods. RESULT: By bacterial culture, the pathogen detection rate from middle ear effusion was 3.6%,while the nasopharynx was 54.0%, the detection rate of Streptococcus pneumonia, Haemophilus influenza, Moraxella catarrhalis was 10.8%, 27.0%, 4.5%, respectively, the drug susceptibility results for 51 samples of bacterial culture positive showed that 39 cases was sensitivite to the β-lactam antibiotic; By PCR, the number of detecting various kinds of bacteria simultaneously in middle ear effusion or in the nasopharynx were 6 and 34. The bacteria prevalences of S. pneumoniae, H. influenzae, M. catarrhalis, and A. otitidis are 5.4%, 5.4%, 3.6%, and 42.3% in the middle ear effusion, are 25.2%, 27.0%,13.5% and 34.2% in nasopharyngeal, respectively. CONCLUSION (1) PCR method is more sensitively detecting the bacteria than conventional bacterial culture methods. (2) The chronic SOM of children may be a combination of mixed bacterial infection, A. otitidis may be the most common pathogen of children SOM. (3) For children of SOM, if antibiotics are chosen to be used early in the disease, we suggest using the β-lactam antibiotics.
Bacteria ; Bacterial Infections ; complications ; Child ; Haemophilus influenzae ; isolation & purification ; Humans ; Moraxella (Branhamella) catarrhalis ; isolation & purification ; Nasopharynx ; Otitis Media with Effusion ; complications ; microbiology ; Polymerase Chain Reaction ; Prevalence ; Streptococcus pneumoniae ; isolation & purification

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

We investigated serotype distribution and antimicrobial resistance of pneumococcal carriage isolates from children after optional immunization with the 7-valent pneumococcal conjugate vaccine (PCV7) in Korea. From June 2009 to June 2010, 205 (16.5%) pneumococcal isolates were obtained from 1,243 nasopharyngeal aspirates of infants and children at Seoul National University Children's Hospital, Korea. Serotype was determined by Quellung reaction and antibiotic susceptibility was tested by E-test. The results were compared to previous studies done in the pre-PCV7 period. In this study, the most common serotypes were 6A (15.3%), 19A (14.7%), 19F (10.2%), 35B (7.3%), and 6D (5.6%). The proportion of PCV7 serotypes decreased from 61.9% to 23.8% (P < 0.001). The overall penicillin nonsusceptibility rate increased from 83.5% to 95.4% (P = 0.001). This study demonstrates the impact of optional PCV7 vaccination in Korea; the proportion of all PCV7 serotypes except 19F decreased while antimicrobial resistant serotypes 6A and 19A further increased.
Anti-Bacterial Agents/pharmacology ; Child, Preschool ; Drug Resistance, Bacterial/drug effects ; Humans ; Infant ; Microbial Sensitivity Tests ; Nasopharynx/*microbiology ; Pneumococcal Infections/immunology/prevention & control ; Republic of Korea ; Serotyping ; Streptococcus pneumoniae/classification/*isolation & purification ; Vaccination ; Vaccines, Conjugate/*immunology

OBJECTIVETo establish multiplex PCR-based assays for detecting H.influenzae and H.parainfluenzae. And the PCR-based assays were applied to detect the carriage rates of H.influenzae and H.parainfluenzae in nasopharyngeal swab specimens which were collected from healthy children.

METHODSMultiplex primers for species-specific PCR were designed by using DNAstar soft based on the sequences of 16S rRNA genes from genus Haemophilus to detect H.influenzae and H.parainfluenzae.

RESULTSThe sensitivity of the 16S rRNA PCR assay for detecting H.influenzae and H.parainfluenzae was 97.53% and 100% respectively, and the specificity was 95.89% and 96.63% respectively. Youden's Index on the ability to detect H.influenzae and H.parainfluenzae was 0.9342 and 0.9663 respectively. 666 nasopharyngeal swab specimens were collected from healthy children. The detection rates of H.influenzae and H.parainfluenzae were 14.11% and 16.07% respectively by using isolation and culture methods. The detection rates of H.influenzae and H.parainfluenzae were 43.54% and 57.96% respectively by 16S rRNA PCR assays. The carriage rates of serotypes a, b, c, d, e, f and non-typeable isolates were 0% (0/666), 0.15% (1/666), 1.20% (8/666), 0.15% (1/666), 1.20% (8/666), 1.80% (12/666), 95.50% (636/666) respectively.

CONCLUSIONThe multiplex PCR assays were very rapid, reliable and feasible methods for detection of H.influenzae and H.parainfluenzae in pharyngeal swab specimens which were compared to conventional isolation and culture methods. 95.5% of H.influenzae strains in healthy children were nontypeable. The encapsulated or typable strains were mainly three serotypes which was c, e, and f serotype.

Haemophilus influenzae ; classification ; genetics ; isolation & purification ; Haemophilus parainfluenzae ; classification ; genetics ; isolation & purification ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Nasopharynx ; microbiology ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Sensitivity and Specificity

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