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Respiratory viruses detection using multiplex PCR in children with severe community acquired pneumonia at Ramathibodi Hospital, Thailand
Oral Presentations / Paediatric Respiratory Reviews 13S1 (2012) S1–S50
ASMC IL-13-induced proliferation and ASMC migration in vitro and therefore may be a new therapeutic option in asthma. F09-13-120 Predictors of postoperative pulmonary complications in children undergoing cardiothoracic surgery M.N. Banque, M.D. Requiron-Sy, M.N. Atienza-De Leon, SalvaniBautista. Division of Pulmonary and Critical Care Medicine, Section of Pediatric Pulmonology, East Avenue, Diliman, Quezon City, Philippines Keywords: Postoperative pulmonary complications, lower pediatric age group, cardiothoracic surgery, congenital heart disease
Background: Postoperative pulmonary complications (PPCs) following cardiothoracic surgery in the lower pediatric age group can never be undermined. Its incidence remained to be alarmingly high. This study was done to determine the risk factors associated with postoperative pulmonary complications following cardiothoracic surgery in children 6 years old and below who underwent cardiothoracic surgery from November 1, 2010 to October 31, 2011 at the Philippine Heart Center. Methods: A prospective cohort study was done among 120 patients. Thirty five risk factors were included including tidal breathing analysis. For qualitative data, proportions/percentages were computed. For quantitative data, means and standard deviation were computed. Comparisons of categories between with and without postoperative complications were done using Chisquare and independent T-test for all continuous variables. All independent variables were entered into a binary logistic regression model. Results: The incidence of PPCs in this study is 73% with atelectasis, pneumonia and air leaks topping the list. The following risk factors were considered to be associated with PPCs: history of respiratory tract infection (RTI), preoperative intubation/mechanical ventilation, high pulmonary artery pressure (PAP), hypercarbia, hemoconcentration, lymphocytopenia, prolonged protime, hypoalbuminemia, high ASA and RACHS-1 scores, prolonged postoperative mechanical ventilation, recovery room (RR) stay, pediatric intensive care unit (PICU) stay and length of postoperative hospital stay. Logistic regression analysis however, only showed history of RTI, preoperative intubation/mechanical ventilation and prolonged postoperative mechanical ventilation as risk factors associated with PPCs. Tidal breathing analysis was not found to be significantly associated with PPCs. Furthermore, as the number of PPCs increases per patient, the higher the risk of mortality. Conclusion: The history of RTI, preoperative mechanical ventilation and prolonged postoperative mechanical ventilation predict postoperative pulmonary complications in children 6 years old and below. With the incidence of PPCs remaining to be high especially in the lower pediatric age group, knowing the risk factors of its occurrence is of paramount importance. F10-13-128 Genotypic, genetic and molecular characteristics of erythromycin C resistant Streptococcus pneumoniae from pediatric patients in Beijing, 2010 L. Zhou, W. Gao, K.H. Yao, A.D. Shen, S.J. Yu, Y.Y. Yang. Beijing Pediatric Research Institute, Beijing, China Background: Streptococcus pneumoniae is a major cause of childhood invasive infections, including sepsis, meningitis and bacteremia, and of non-invasive disease such as otitis media, bronchitis and pneumonia. Antibiotics are often the first choice for pneumococcal infections. However, S. pneumoniae is difficult to treat during recent years because of increasing resistance to various antibiotics, including macrolides and tetracyclines. In S. pneumoniae, most macrolide-resistant strains are also resistant to tetracycline. The frequent association of erythromycin and tetracycline resistance is due to the insertion of the ermB gene into conjugative and composite transposons of the Tn916 or Tn917 family (Tn6002,
S49
Tn3872, Tn1545, and Tn6003) which harbor the tet(M) gene. The aim of this study was to detect the resistance of S. pneumoniae to erythromycin and tetracycline collected from pediatric patients with upper respiratory infections in Beijing, 2010, and to analyze the phenotypic and genotypic characteristics of erythromycin-resistant pneumococcal isolates as well as the molecular epidemiology. Methods: 140 isolates of S. pneumoniae were collected at Beijing Children’s Hospital. E-test was applied to detect the resistance of these isolates to erythromycin and tetracycline. Macrolide resistance phenotype was performed by the double-disk diffusion method, and the macrolide resistance genes ermB and mef were detected by polymerase chain reaction (PCR) while the Tn916 and Tn917 transposon-related genes (int, xis, tnpA, and tnpR), the tetracycline resistance gene tetM and the promoter of the aph3 -III gene were also detected by PCR. House-keeping genes aroE, gdh, gki, recP, spi, xpt, and ddl were amplified by PCR. The sequences at each of the 7 loci were then compared with the sequences of all of the known alleles at those loci in the database at the pneumococcal MLST website as well as sequence types (STs). The eBURST v3 software was used to investigate the affinity relationships among the isolates and to assign strains to a clone complex when using the stringent group definition of six of seven shared alleles. Results: The resistance rate of 140 S. pneumoniae isolates to erythromycin was 96.4% while it was 79.3% to tetracycline. Among the 135 erythromycin-resistant strains, 91.1% of isolates were nonsusceptible (intermediate and resistant) to tetracycline and most isolates had cMLSB phenotype. The ermB and tetM genes were the main mechanisms for erythromycin and tetracycline resistance, respectively. All erythromycin-resistant isolates were tested by PCR for the presence of the int, xis, tnpA, and tnpR genes associated with conjugative transposons (Tn916 or Tn917 family). Tn6002, positive detection of ermB, tetM, int, and xis genes, was the most common transposon for both erythromycin and tetracycline resistance. A total of 134 erythromycin-resistant S. pneumoniae isolates (one isolate died during keeping) were done for MLST analysis. The prevailing sequence types were ST271, ST81, and ST876 while CC271 was the most frequent clone complex, accounting for 24.6% among the erythromycin-resistant pneumococci. Conclusions: The majority of erythromycin-resistant pneumococcal isolated in China had the MLSB phenotype, and carried the ermB, tetM, xis, and int genes, suggesting the spread of transposons of the Tn916 family. Further long-term surveys are required to monitor antibiotic resistance, phenotype, genotype, and sequence type in S. pneumoniae. F11-13-242 Respiratory viruses detection using multiplex PCR in children with severe community acquired pneumonia at Ramathibodi Hospital, Thailand T. Wannachai, H. Kamalaporn, A. Preutthipan. Ramathibodi Hospital, Mahidol University Pediatric Pulmonology, Bangkok, Thailand Background: Most of community acquired pneumonia (CAP) in children derived from viral etiology. The epidemiology and characteristics of viral associated CAP in Thai children remain poorly defined. Since the novel multiplex PCR assay has recently been developed, as many as 18 respiratory viruses can be detected from nasopharyngeal specimens. Objectives: To determine the viral epidemiology and characteristics of severe CAP in children at Ramathibodi hospital Methods: An 8-month prospective study was conducted during April to November 2011. Children younger than 18 years old with severe CAP according to WHO criteria were recruited. Nasopharyngeal specimens were obtained within 48 hours after admission. Viral detection was done using multiplex PCR to detect 18 respiratory viruses including adenovirus, rhinovirus/enterovirus, 4 human coronaviruses, 3 influenza viruses type A, influenza virus type B, 4 parainfluenza viruses, respiratory syncytial virus
S50
Oral Presentations / Paediatric Respiratory Reviews 13S1 (2012) S1–S50
A and B, bocavirus and human metapneumovirus. The patients were categorized into rhinovirus and non-rhino virus infection groups. Demographic data, etiologic agents and outcome were compared between the two groups using chi-square and Student t-tests. Results: During 8-month period, there were 53/77(68.8%) children with severe CAP who had positive viral PCR results. The median age was 57.4 (range 2 months to 17 years). Thirty-six (67%) were male. The peak prevalence was in August (n = 16; 30%). Twenty patients (35.8%) required non-invasive positive pressure ventilation, whereas 19 (37.7%) required mechanical ventilation. The most common viruses found were rhinovirus (n = 30; 56.6%), human metapneumovirus (n = 8; 15.1%), respiratory syncytial virusA (n = 6; 11.3%). Six patients (11.3%) had multiple viral infections. Thirteen patients (24.5%) had positive sputum culture for bacteria. There were no statistically significant differences in demographic data, etiologic agents or underlying diseases between rhino virus and non-rhinovirus infection group. Wheezing was found in half of the patients with severe viral CAP (29 of 53, 54.7%). Seventeen of 29 (58.6%) had rhinovirus positive, 9 of these are the first wheezers. Ninety percent (27 of 30) of patients with positive rhinovirus required ventilator support which is much more than in patients with hMPV (5 of 8, 62.5%) and RSV (3 of 7, 42.9%). Rhinovirus infections are present throughout the year in every age group and the peak of infection is mainly in the rainy season. Human metapneumovirus are the second most common in children under five. No patients died. Conclusion: By using multiplex PCR to detect 18 viral pathogens, rhinovirus is the most common viral agents identified in severe CAP in children at our institution. Patients with positive rhinovirus may present with wheeze and had more chance to require ventilatory support.
ASMC IL-13-induced proliferation and ASMC migration in vitro and therefore may be a new therapeutic option in asthma. F09-13-120 Predictors of postoperative pulmonary complications in children undergoing cardiothoracic surgery M.N. Banque, M.D. Requiron-Sy, M.N. Atienza-De Leon, SalvaniBautista. Division of Pulmonary and Critical Care Medicine, Section of Pediatric Pulmonology, East Avenue, Diliman, Quezon City, Philippines Keywords: Postoperative pulmonary complications, lower pediatric age group, cardiothoracic surgery, congenital heart disease
Background: Postoperative pulmonary complications (PPCs) following cardiothoracic surgery in the lower pediatric age group can never be undermined. Its incidence remained to be alarmingly high. This study was done to determine the risk factors associated with postoperative pulmonary complications following cardiothoracic surgery in children 6 years old and below who underwent cardiothoracic surgery from November 1, 2010 to October 31, 2011 at the Philippine Heart Center. Methods: A prospective cohort study was done among 120 patients. Thirty five risk factors were included including tidal breathing analysis. For qualitative data, proportions/percentages were computed. For quantitative data, means and standard deviation were computed. Comparisons of categories between with and without postoperative complications were done using Chisquare and independent T-test for all continuous variables. All independent variables were entered into a binary logistic regression model. Results: The incidence of PPCs in this study is 73% with atelectasis, pneumonia and air leaks topping the list. The following risk factors were considered to be associated with PPCs: history of respiratory tract infection (RTI), preoperative intubation/mechanical ventilation, high pulmonary artery pressure (PAP), hypercarbia, hemoconcentration, lymphocytopenia, prolonged protime, hypoalbuminemia, high ASA and RACHS-1 scores, prolonged postoperative mechanical ventilation, recovery room (RR) stay, pediatric intensive care unit (PICU) stay and length of postoperative hospital stay. Logistic regression analysis however, only showed history of RTI, preoperative intubation/mechanical ventilation and prolonged postoperative mechanical ventilation as risk factors associated with PPCs. Tidal breathing analysis was not found to be significantly associated with PPCs. Furthermore, as the number of PPCs increases per patient, the higher the risk of mortality. Conclusion: The history of RTI, preoperative mechanical ventilation and prolonged postoperative mechanical ventilation predict postoperative pulmonary complications in children 6 years old and below. With the incidence of PPCs remaining to be high especially in the lower pediatric age group, knowing the risk factors of its occurrence is of paramount importance. F10-13-128 Genotypic, genetic and molecular characteristics of erythromycin C resistant Streptococcus pneumoniae from pediatric patients in Beijing, 2010 L. Zhou, W. Gao, K.H. Yao, A.D. Shen, S.J. Yu, Y.Y. Yang. Beijing Pediatric Research Institute, Beijing, China Background: Streptococcus pneumoniae is a major cause of childhood invasive infections, including sepsis, meningitis and bacteremia, and of non-invasive disease such as otitis media, bronchitis and pneumonia. Antibiotics are often the first choice for pneumococcal infections. However, S. pneumoniae is difficult to treat during recent years because of increasing resistance to various antibiotics, including macrolides and tetracyclines. In S. pneumoniae, most macrolide-resistant strains are also resistant to tetracycline. The frequent association of erythromycin and tetracycline resistance is due to the insertion of the ermB gene into conjugative and composite transposons of the Tn916 or Tn917 family (Tn6002,
S49
Tn3872, Tn1545, and Tn6003) which harbor the tet(M) gene. The aim of this study was to detect the resistance of S. pneumoniae to erythromycin and tetracycline collected from pediatric patients with upper respiratory infections in Beijing, 2010, and to analyze the phenotypic and genotypic characteristics of erythromycin-resistant pneumococcal isolates as well as the molecular epidemiology. Methods: 140 isolates of S. pneumoniae were collected at Beijing Children’s Hospital. E-test was applied to detect the resistance of these isolates to erythromycin and tetracycline. Macrolide resistance phenotype was performed by the double-disk diffusion method, and the macrolide resistance genes ermB and mef were detected by polymerase chain reaction (PCR) while the Tn916 and Tn917 transposon-related genes (int, xis, tnpA, and tnpR), the tetracycline resistance gene tetM and the promoter of the aph3 -III gene were also detected by PCR. House-keeping genes aroE, gdh, gki, recP, spi, xpt, and ddl were amplified by PCR. The sequences at each of the 7 loci were then compared with the sequences of all of the known alleles at those loci in the database at the pneumococcal MLST website as well as sequence types (STs). The eBURST v3 software was used to investigate the affinity relationships among the isolates and to assign strains to a clone complex when using the stringent group definition of six of seven shared alleles. Results: The resistance rate of 140 S. pneumoniae isolates to erythromycin was 96.4% while it was 79.3% to tetracycline. Among the 135 erythromycin-resistant strains, 91.1% of isolates were nonsusceptible (intermediate and resistant) to tetracycline and most isolates had cMLSB phenotype. The ermB and tetM genes were the main mechanisms for erythromycin and tetracycline resistance, respectively. All erythromycin-resistant isolates were tested by PCR for the presence of the int, xis, tnpA, and tnpR genes associated with conjugative transposons (Tn916 or Tn917 family). Tn6002, positive detection of ermB, tetM, int, and xis genes, was the most common transposon for both erythromycin and tetracycline resistance. A total of 134 erythromycin-resistant S. pneumoniae isolates (one isolate died during keeping) were done for MLST analysis. The prevailing sequence types were ST271, ST81, and ST876 while CC271 was the most frequent clone complex, accounting for 24.6% among the erythromycin-resistant pneumococci. Conclusions: The majority of erythromycin-resistant pneumococcal isolated in China had the MLSB phenotype, and carried the ermB, tetM, xis, and int genes, suggesting the spread of transposons of the Tn916 family. Further long-term surveys are required to monitor antibiotic resistance, phenotype, genotype, and sequence type in S. pneumoniae. F11-13-242 Respiratory viruses detection using multiplex PCR in children with severe community acquired pneumonia at Ramathibodi Hospital, Thailand T. Wannachai, H. Kamalaporn, A. Preutthipan. Ramathibodi Hospital, Mahidol University Pediatric Pulmonology, Bangkok, Thailand Background: Most of community acquired pneumonia (CAP) in children derived from viral etiology. The epidemiology and characteristics of viral associated CAP in Thai children remain poorly defined. Since the novel multiplex PCR assay has recently been developed, as many as 18 respiratory viruses can be detected from nasopharyngeal specimens. Objectives: To determine the viral epidemiology and characteristics of severe CAP in children at Ramathibodi hospital Methods: An 8-month prospective study was conducted during April to November 2011. Children younger than 18 years old with severe CAP according to WHO criteria were recruited. Nasopharyngeal specimens were obtained within 48 hours after admission. Viral detection was done using multiplex PCR to detect 18 respiratory viruses including adenovirus, rhinovirus/enterovirus, 4 human coronaviruses, 3 influenza viruses type A, influenza virus type B, 4 parainfluenza viruses, respiratory syncytial virus
S50
Oral Presentations / Paediatric Respiratory Reviews 13S1 (2012) S1–S50
A and B, bocavirus and human metapneumovirus. The patients were categorized into rhinovirus and non-rhino virus infection groups. Demographic data, etiologic agents and outcome were compared between the two groups using chi-square and Student t-tests. Results: During 8-month period, there were 53/77(68.8%) children with severe CAP who had positive viral PCR results. The median age was 57.4 (range 2 months to 17 years). Thirty-six (67%) were male. The peak prevalence was in August (n = 16; 30%). Twenty patients (35.8%) required non-invasive positive pressure ventilation, whereas 19 (37.7%) required mechanical ventilation. The most common viruses found were rhinovirus (n = 30; 56.6%), human metapneumovirus (n = 8; 15.1%), respiratory syncytial virusA (n = 6; 11.3%). Six patients (11.3%) had multiple viral infections. Thirteen patients (24.5%) had positive sputum culture for bacteria. There were no statistically significant differences in demographic data, etiologic agents or underlying diseases between rhino virus and non-rhinovirus infection group. Wheezing was found in half of the patients with severe viral CAP (29 of 53, 54.7%). Seventeen of 29 (58.6%) had rhinovirus positive, 9 of these are the first wheezers. Ninety percent (27 of 30) of patients with positive rhinovirus required ventilator support which is much more than in patients with hMPV (5 of 8, 62.5%) and RSV (3 of 7, 42.9%). Rhinovirus infections are present throughout the year in every age group and the peak of infection is mainly in the rainy season. Human metapneumovirus are the second most common in children under five. No patients died. Conclusion: By using multiplex PCR to detect 18 viral pathogens, rhinovirus is the most common viral agents identified in severe CAP in children at our institution. Patients with positive rhinovirus may present with wheeze and had more chance to require ventilatory support.