Kim, Su-Nam;Min, Kwan-Ki;Kim, Seung-Hwan;Choi, In-Hwa;Lee, Suhk-Hyung;Pyo, Suhk-Noung;Rhee, Dong-Kwon
Journal of Microbiology
/
v.34
no.2
/
pp.179-183
/
1996
Streptoccus Pneumoniae (pneumococcus), the most common cause of bacterial pneumonia, has an ample polysaccharide (PS) capsule that is highly antigenic and is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsulard PS by type 1 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based media were found to support luxuriant growth of pneumococcus type 1 at the same level. Because BHI medium is rather expensive and more complex than the Casitone based media, the Casitone based media was uwed to study optimization of the culture condition. The phase of growth which accomodated maximum PS production was logarithmic phase. Concentrations of glucose greater than 0.2% did not ehnahce growth or PS production. Substitution of netrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium ion) had adverse affects on growth and PS production. On the other hand, low level aeration was beneficial for increased PS production. Addition of 3 mg/1 concentration of methionine, phenylalanine, and threonine were found to enhance growth and PS production. The synerigistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cummulative increase in capsular PS production.
Purpose : Streptococcus pneumoniae remains a leading cause of meningitis, sepsis, pneumonia, and otitis media in children worldwide. Emergence of drug-resistant organism has substantially complicated the therapy of these infections. This study was conducted to determine the clinical features and changing patterns of antibiotic-resistant rates of Streptococcus pneumoniae. Methods : We have retrospectively examined 306 cases of culture proven patients who were admitted to the Department of Pediatrics, Severance hospital, from the year 1991 to year 2000. The type of culture specimen used, presence of penicillin resistance, characteristics according to various presenting diseases and their prevailing year were also examined. Results : The mean age of cases was 7.9 year and the ratio of male to female was 1.6 : 1. The main age group of pneumococcal infection was under 2 years(42%). Systemic infections associated with pneumococci were sepsis(19.3%) and meningitis(9.5%), while local infections presented as pneumonia(29.2%), otitis media(19.3%), exudative tonsillitis(13.3%), and sinusitis(9.2%) in the order of frequency. Seasonal variation was seen in the incidence of pneumococcal infection: high incidence of infection was seen in Spring(Mar.~Apr; 32%), while the incidence was low during summer(Aug.~Sep.; 6%). Penicillin-resistant rate of pneumococci was steadily on the increase since the year 1991(65%) to year 2000(84%). Conclusion : Antibiotic-resistant pneumococci increased during the past decade. For effective prevention of pneumococcal infections, national survey of pneumococcal infections and expanded use of pneumococcal vaccination would be needed.
Byun, Sung Wan;Kim, Han Wool;Yoon, Seo Hee;Park, In Ho;Kim, Kyung-Hyo
Pediatric Infection and Vaccine
/
v.22
no.2
/
pp.106-112
/
2015
Purpose: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. Methods: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. Results: The detection limit of the PCR assay was approximately $10^4$ colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over $10^6$ CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). Conclusions: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA -specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.
Lee, Hyeon Seung;Choe, Young June;Cho, Eun Young;Lee, Hyunju;Choi, Eun Hwa;Lee, Hoan Jong
Pediatric Infection and Vaccine
/
v.21
no.3
/
pp.207-213
/
2014
Purpose: This study aimed to investigate the association between respiratory virus infection and pneumococcal colonization in children. Methods: From May 2009 to June 2010, nasopharyngeal (NP) aspirates were obtained from patients under 18 years old who visited Seoul National University Children's Hospital for respiratory symptoms. NP samples were used to detect respiratory viruses (influenza virus A and B, parainfluenza virus 1, 2 and 3, respiratory syncytial virus A and B, adenovirus, rhinovirus A/B, human metapneumovirus, human coronavirus 229E/NL63 and OC43/HKU1) by RT-PCR and pneumococcus by culture. Results: Median age of the patients was 27 months old. A total of 1,367 NP aspirates were tested for respiratory viruses and pneumococcus. Pneumococcus was isolated from 228 (16.7%) of samples and respiratory viruses were detected from 731 (53.5%). Common viruses were rhinovirus (18.4%), respiratory syncytial virus (RSV) A (10.6%), adenovirus (6.9%), influenza virus A (6.8%). Pneumococcal isolation rate was significantly higher in the cases of positive virus detection than negative detection [21.3% (156/731) vs. 11.3% (72/636), P <0.001]. For individual viruses, pneumococcal isolation rate was positively associated with detection of influenza virus A [24.7% (23/93) vs 16.1% (205/1274), P=0.001], RSV A [28.3% (41/145) vs 15.3% (187/1222), P=0.001], RSV B [31.3% (10/32) vs 16.3% (218/1335), P=0.042], rhinovirus A/B [22.6% (57/252) vs 15.3% (171/1115), P=0.010]. Conclusion: The study revealed that pneumococcal isolation from NP aspirates is related with respiratory virus detection. The result of this study could be used to investigate how respiratory viruses and pneumococcus cause clinical diseases.
Jung, Jiwon;Yoo, Ree Nar;Sung, Hungseop;Kim, Mina;Lee, Jina
Pediatric Infection and Vaccine
/
v.26
no.1
/
pp.1-10
/
2019
Purpose: We investigated the distribution and antimicrobial resistance of pneumococcal isolates from hospitalized children at Asan Medical Center for recent 4 years, and aimed to recommend proper choice of empirical antibiotics for pneumococcal infection. Methods: From March 2014 to May 2018, children admitted to Asan Medical Center Childrens' Hospital with pneumococcal infection were subjected for evaluation of minimal inhibitory concentration (MIC) for ${\beta}-lactams$ and macrolide antibiotics. Patient's age, underlying disease, gender were retrospectively collected. Using Monte Carlo simulation model and MIC from our study, we predicted the rate of treatment success with amoxicillin treatment. Results: Sixty-three isolates were analyzed including 20.6% (n=13) of invasive isolates, and 79.4% (n=50) of non-invasive isolates; median age were 3.3 years old, and 87.3% of the pneumococcal infections occurred to children with underlying disease. Overall susceptibility rate was 49.2%, 68.2%, and 74.6% for amoxicillin, parenteral penicillin, and cefotaxime respectively. 23.8% and 9.5% of the isolates showed high resistance for amoxicillin, and cefotaxime. Only 4.8% (n=3) were susceptible to erythromycin. Monte Carlo simulation model revealed the likelihood of treatment success was 46.0% at the dosage of 90 mg/kg/day of amoxicillin. Conclusions: Recent pneumococcal isolates from pediatric patients with underlying disease revealed high resistance for amoxicillin and cefotaxime, and high resistance for erythromycin. Prudent choice of antibiotics based on the local data of resistance cannot be emphasized enough, especially in high risk patients with underlying disease, and timely vaccination should be implemented for prevention of the spread of resistant strains.
Most boys afflicted with panhypogammaglobulinemia, also known as X-linked agammaglobulinemia, remain healthy during the first 6 to 12 months of life because of protection by maternally transmitted IgG antibodies. Thereafter, they repeatedly acquire infections with high-grade pathogens, such as pneumococci, streptococci, and hemophilus unless given antibiotics or immunoglobulin replacement therapy. We experienced a case of panhypogammaglobulinemia in a 4 years old boy. He had been suffered from recurrent upper respiratory tract infection, otitis media and pneumonia since late infancy. He was admitted due to right pleural effusion with pneumonia, and streptococcus pneumoniae was isolated from pleural fluid and blood cultures. His immune status revealed panhypogammaglobulinemia and deficiency in mature B lymphocyte. He was treated with appropriate antibiotics therapy, but showed poor responses. He was transferred to department of thoracic surgery, and received minithoracotomy (decortication) operation. He was successfully treated with operation, antibiotics, and IV gammaglobulin infusions. Now he is being followed with periodic IV gammaglobulin replacement therapy.
Purpose : To characterize the epidemiology and clinical features of invasive pneumococcal infections in Korean children. Methods : One hundred ninety four cases of invasive pneumococcal infections diagnosed at the Seoul National University Children's Hospital from October 1985 to December 2003 were analysed retrospectively. All isolates were screened for resistance to penicillin by oxacillin disc diffusion test. Serotypes were determined for 125 isolates. Results : The types of infection were bacteremia without focus 84/194(43%), meningitis 36/194(19%), pneumonia with bacteremia 36/194(19%), peritonitis 24/194(12%), other focal infections 3/194(2%). Fifty seven percent(110/194) of the episodes developed in the immunocompromised and 20%(37/194) were nosocomially acquired. The patients younger than 2 years of age was 60% in the immunocompetent patients and 25% in the immunocompromised patients. The overall case fatality rate was 7%. All the isolates by 1988 were susceptible to penicillin screened by oxacillin disk. Penicillin resistance was first detected in 1989(20%), and then increased rapidly; 89% in 1995, 69% in 1996, and 80~100% thereafter. The seven most frequently isolated serotypes were 23F, 19F, 14, 6B, 6A, 9V and 19A, which accounted for 70% of total isolates. Conclusion : S. pneumoniaeis an important cause of morbidity and mortality in children. Invasive infections caused by S. pneumoniae most often occurred in infants and young children, while they are frequent in older immunocompromised children as well. This is the largest case series on invasive pneumococcal infections in Korean children.
Kim, Young Kwang;Lee, Kyoung Hoon;Yun, Ki Wook;Lee, Mi Kyung;Lim, In Seok
Pediatric Infection and Vaccine
/
v.23
no.3
/
pp.194-201
/
2016
Purpose: Monitoring pneumococcal carriage rates is important. We developed and evaluated the accuracy of a real-time reverse transcription polymerase chain reaction (RT-PCR) protocol for the detection of Streptococcus pneumoniae. Methods: In October 2014, 157 nasopharyngeal aspirates were collected from patients aged <18 years admitted to Chung-Ang University Hospital. We developed and evaluated a real-time PCR method for detecting S. pneumoniae by comparing culture findings with the results of the real-time PCR using genomic DNA (gDNA). Of 157 samples, 20 specimens were analyzed in order to compare the results of cultures, real-time PCR, and real-time RT-PCR. Results: The concordance rate between culture findings and the results of real-time PCR was 0.922 (P<0.01, Fisher exact test). The 133 culture-negative samples were confirmed to be negative for S. pneumoniae using real-time PCR. Of the remaining 24 culture-positive samples, 21 were identified as S. pneumonia -positive using real-time PCR. The results of real-time RT-PCR and real-time PCR from 20 specimens were consistent with culture findings for all S. pneumoniae -positive samples except one. Culture and real-time RT-PCR required 26.5 and 4.5 hours to perform, respectively. Conclusions: This study established a real-time RT-PCR method for the detection of pneumococcal carriage in the nasopharynx. Real-time RT-PCR is an accurate, convenient, and time-saving method; therefore, it may be useful for collecting epidemiologic data regarding pneumococcal carriage in children.
Purpose : Pneumoccocus is one of the most important causes of invasive infection through the childhood period and the prevelance of antibiotics resistance of pneumococcus is increasing worldwide. A 7-valent conjugate vaccine has been developed. It is important to know the prevalence of each serotype of pneumococci in the countries where the vaccine is used to estimate the coverage rate by the vaccine. Methods : One hundred and twenty seven strains of clinical isolates and 72 strains from healthy carriers recovered from Korean children during the period from 1997 to 2002 were subjected to determination of serotype by Quellung reaction and penicillin susceptibility with oxacillin disc diffusion test. Results : Forty-three per cent of clinical isolates were obtained from children under two years of age. Thirty strains(24%) were isolated from normally sterile body fluids. The frequent serotypes were 19F, 19A, 23F, 6A, 6B and 9V. Fifty-six per cent of the clinical isolates were represented in the current 7-valent protein conjugate pneumococccal vaccine, and 84% when the cross-reactive serotypes were included. Frequent serotypes of strains isolated from one to five year-old healthy children were 19F, 14, 11A, 23F, 18C, and 19A. Seventy-one per cent of the carrier strains were included in the 7-valent vaccine. Ninety-three per cent of the clinical isolates and 86% of carrier strains were not susceptible to penicilline. Conclusion : Fifty-six to 84% of pneumococci recovered from Korean children are covered by the current 7-valent protein conjugate pneumococcal vaccine and the prevalence of penicillin resistance was very high.
Chung, Eun Hee;Ma, Sang Hyuck;Hong, Young Jin;Kim, Kyung Hyo;Kim, Jong-Hyun;Lee, Jin A;Lee, Hoan Jong
Pediatric Infection and Vaccine
/
v.13
no.2
/
pp.163-173
/
2006
Purpose : To reduce the number of injections necessary to vaccinate young infants, various combined vaccines have been developed. The $Comvax^{TM}$ manufactured by Merck & Co. is a combination of Hepatitis B and PRP-OMP conjugate Haemophilus influenzae Type b vaccine. The purpose of this study is to evaluate the immunogenicity and safety of $Comvax^{TM}$ in Korean infants. Methods : The infants who were vaccinated at 0 months of age with Hepatitis B vaccine, were recruited for this study after parental informed consent was obtained. The subjects were vaccinated with $Comvax^{TM}$ at 2 and 4 months of age. At each visit, infants were also immunized with DTaP, inactivated poliovirus vaccine, and pneumococcal vaccine when indicated. The serum anti-PRP and anti-HBs were measured at 2 months after the 1st dose(4 months age), and the 2nd dose(6 months age) by the ELISA and chemiluminescent microparticle immunoassay method, respectively. The local and systemic adverse reactions of vaccination were monitored for 3 consecutive days after each immunization. Results : Among sixty-five healthy infants(35 male infants) enrolled in this study; fifty eight(32 male infants) completed the scheduled immunizations. The geometric mean titers (GMTs) of anti-PRP at 2 months after the 1st dose and the 2nd dose were 1.96 ${\mu}g/mL$ (95% CI; 1.38~2.78) and 10.02 ${\mu}g/mL$ (95% CI; 7.04~14.26), respectively. Anti-PRP ${\geq}1.0$${\mu}g/mL$, was obtained in 63.2%(95% CI; 53.75~72.65) after 1 dose, and 96.6%(95% CI; 93.05~100) after 2 doses. The GMTs of anti-HBs were 38.32 mIU/mL(95% CI; 22.42~65.51), and 101.17 mIU/mL(95% CI; 65.94~155.25) at 2 month after the 1st dose and 2nd dose of $Comvax^{TM}$, respectively. Anti-HBs ${\geq}10$ mIU/mL was observed in 73.7%(95% CI; 65.07~82.33) after 1 dose and 94.8%(95% CI; 90.45~99.15) after 2 doses. Most of the adverse reactions after vaccination were mild. Irritability, the most common systemic reaction, was observed in 24.8%, followed by drowsiness(19.2%), poor feeding(19.2%) and fever(7.2%). Among the local reactions tenderness was observed in 25.6%, redness(${\geq}5$ mm) in 19.2% and swelling(${\geq}5$ mm) in 4.8%. Conclusion : The $Comvax^{TM}$ vaccine was highly immunogenic for PRP and safe in Korean infants. Although the hepatitis B vaccine component was administered at 0, 2, 4 months, this study showed good immunogenicity against HBsAg.
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