• Korean Journal of Microbiology
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• v.38 no.1
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• pp.38-44
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• 2002

Pneumococcal surfacce protein A (PspA) is an important virulence factor and an antigenically variable surface protein of the pneumococci. To purify the PspA from S. pneumoniae KNIH1156 , a clinical isolate (type 19F), we have taken advantage of the fact that PspA is released from the surface of pneumococci into the medium by growing in a CDM-ET medium and PspA is capable of binding human lactoferrin, the iron carrier protein. PspA of S. pneumoniae KNIH1156 was purified from culture supernatant by human lactoferrin (hLf) affinity chromatography. The purified PspA was confirmed with anti-PspA antiserum and also had the binding capacity to hLf specifically. To determine whether the purified PspA could elicit protection in mice against pneumococcal inflection, we immunized the mice with purified PspA and subsequently challenged with S. pneumoniae KNIH1156. Immunization with purified PspA protected mice from 500 times the $LD^{50}$ of S. pneumoniae KNIH1156. Therefore, it has been shown that purified PspA fromS. pneumoniae KNIH1156 (type 19F) is a protective immunogen.

• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.734-738
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• 2011

Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. There are currently 90 capsular serotypes identified and 23 serotypes (1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F) are responsible for about 90% of invasive disease. Among the more than 90 different S. pneumoniae serotypes, serotype 19A is globally very prevalent. A simplified purification procedure including adjustment of cell lysate pH to 4.5, fractionation with 50. 80% ethanol, and dialysis rendered capsular polysaccharide (CPS) in a yield of $31.32{\pm}3.11$ mg from 1 l culture (75% recovery after lyses). The product contained only 69.6 ${\mu}g$ of protein (99.78% purity) and 0.8mg (sum of the precipitants from 50~60%, 60~70%, and 70~80%) of nucleic acid (97.45% purity). The purified CPS was conjugated with bovine serum albumin; the product size ranged from 100 to 180 kDa.

• Clinical and Experimental Pediatrics
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• v.49 no.3
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• pp.235-241
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• 2006

Streptococus pneumoniae is an important cause of invasive infections as well as non-invasive infections such as acute otitis media and sinusitis both in children and adults. Resistance of S. pneumoniae to multiple antimicrobials is increasing and poses therapeutic challenges, and prevention became more important. 23-valent polysaccharide vaccine has been used for the last several decades, but is not effective in children <2 years of age, the highest risk group of invasive diseases. Recently, a 7-valent pneumococcal protein conjugate vaccine(PCV) which is effective in infants and young children has been developed. The efficacy of PCVs against invasive pneumococcal disease and pneumonia is well established and is documented in several well-conducted studies. However, the effect of PCVs on otitis media is less obvious and more complex. PCVs clearly reduce diseases caused by vaccine-type(VT) pneumococci, but replacement of VT serotypes by non-VT serotypes in nasopharyngeal carriage of S. pneumoniae is responsible for the increase in acute otitis media caused by non-VT serotypes. Three years after introduction of PCV in the US, some increase of invasive infections with serotype 19A possibly due to serotype switching within certain vaccine type strains has been noted. Since most antibiotic-resistance in S. pneumoniae is confined to VT serotypes, vaccine use also reduces antibiotic resistance. With development of PCV, there was a great advance in the prevention of pneumococcal diseases, but replacement with potential virulent organisms and development of antibiotic resistance in non-VT pneumococci is a possibility that needs careful monitoring.

• Archives of Pharmacal Research
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• v.19 no.3
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• pp.173-177
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• 1996

The Pneumococcus, Streptococcus pneumoniae, has an ample polysaccharide (PS) capsule that is highly antigenic and is the main virulence factor of the organism. The capsular PS is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 4 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based medium were found to support luxuriant growth of pneumococcus type 4 at the same level. Therefore in this study, the Casitone based medium was used to study optimization of the culture condition because of BHI broth's high cost and complex nature. The phase of growth which accomodated maximum PS production was exponential phase. Concentrations of glucose greater than 0.8% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium, copper, and magnesium ions) had adverse effects on growth and PS production. On the other hand, low level aeration and supplementation of 3 mg/l concentration of asparagine, phenylalanine, or threonine were beneficial for increased PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

• Pediatric Infection and Vaccine
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• v.13 no.2
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• pp.124-129
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• 2006

Purpose : We investigated the causative organisms of bacterial meningitis by age distribution from 1996 to 2005. Methods : Retrospective data were obtained from the medical records with diagnosis of bacterial meningitis or neonatal meningitis from 1996 through 2005. A case was defined by isolation of organism or detection of its antigen by latex agglutination from cerebrospinal fluid. Results : A total of 46 cases(27 neonates and 19 children) were identified. 15 of 27 episodes(55.6%) of neonatal meningitis had a concomitant-positive blood culture. Group B streptococci were the most common bacterial causes of neonatal meningitis(44.4%). Nine of 12 episodes(75.0%) were late-onset infections in neonatal meningitis caused by group B streptococci. 16 of 19 children(84.2%) with bacterial meningitis beyond the neonatal period were younger than 5 years of age(median age, 23 months). Of 19 cases, 8 infections were with Streptococcus pneumoniae, 8 were with Haemophilus influenzae and 3 were with Neisseria meningitidis. Since 2001 there was no case of meningococcal meningitis in this study. Conclusion : In neonates group B streptococci are the most common causative organisms of bacterial meningitis, especially late-onset infections. In infants and young children, the predominant causes of bacterial meningitis are H. influenzae and S. pneumoniae; meningitis caused by the former are likely to decrease after the introduction of the conjugate vaccine for H. influenzae type b.

• Journal of Microbiology
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• v.41 no.1
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• pp.1-6
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• 2003

The genetic relatedness of multidrug-resistant pneumococcal isolates of serotypes 19F and 23F was investigated. The DNA fragments digested with Sma I were resolved by pulsed-field gel electrophoresis (PFGE). PFGE analysis of 365. pneumoniae isolates showed 13 different patterns. Among 22 isolates of serotype 19F, 9 different PFGE patterns were present and 14 isolates of serotype 23F isolates represented 5 distinct PFGE patterns. Two isolates of serotype 19F and six isolates of serotype 23F shared the same PFGE pattern (Pattern I). Based on the genetic relatedness within the strains (one genetic cluster was defined as having more than 85% homology), we divided the pneumococcal strains into genefic clusters (Ⅰ, II, III, IV, V, and VI). The 22 strains of serotype 19F belonged to five distinct genetic clusters (I, II, III, IV, V and VI) and 14 strains of serotype 23F represented two genetic clusters (I and II ). These results showed that strains of serotype 19F are genetically more diverse than those of serotype 23F, Serotype 19F isolates with PFGE patterns H and I appeared to be less related to those of the remaining PFCE patterns (A to G) (less than 60% genetic relatedness), but those strains were genetically closely related with serotype 23f. These results suggest that the latter isolates originated from horizontal transfer of the capsular type 19F gene locus to 23F pneumococcal genotypes. In conclusion, the multidrug-resistant pneumococcal isolates of serotype 19f and 23F isolated in Korea are the result of the spread of a limited number of resistant clones.

• Clinical and Experimental Pediatrics
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• v.50 no.2
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• pp.157-162
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• 2007

Purpose : The purpose of this study was to analyze the epidemiology, causative organism, clinical manifestation and prognosis of bacterial meningitis for children after the introduction of Haemophilus influenzae type b (Hib) vaccine in Daejeon and Chungcheong area. Methods : We analyzed retrospectively 53 medical records who had been diagnosed with bacterial meningitis at 10 general or university hospitals in Daejeon and Chungcheong area. All patients aged 1 month-14 years admitted between January 2001 through December 2005. Results : During the 5-year study period, 40 of all cases were positive for bacterial growth. Of the 40 cases that were CSF culture-proven bacterial meningitis, Streptococcus peumoniae was the most common bacteria for 17 (32.1%) of all cases, followed by H. influenzae for 10 (18.9%), Neisseria meningitidis for 3 (5.7%). In this study, the most common clinical manifestation is fever, accompanied by all cases. CSF leukocyte count was more than $100/mm^3$ in 45 (84.1%) cases. CSF glucose concentration was less than 50 mg/dL in 42 (79.2%) cases and protein concentration was more than 45 mg/dL in 49 (92.5%) cases. 45 of all cases made a recovery after treatment and were discharged. Most common complication after treatment is subdural effusion (19.0%) and hearing disturbance (9.4%). Conclusion : The most common organism of culture-proven bacterial meningitis in the children beyond neonatal period was S. pneumoniae. Continued surveillance studies were demanded to know the altered incidence of bacterial meningitis, because we expect the incidence of S. pneumoniae meninigitis will be on the decrease after more active innoculation of pneumococcal protein conjugate vaccine.

• Clinical and Experimental Pediatrics
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• v.46 no.4
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• pp.345-350
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• 2003

Purpose : Bacterial meningitis is a serious infection of childhood associated with a significant morbidity and mortality. Repeated cerebrospinal fluid(CSF) examination is a useful prognostic indicator and a delayed sterilization is associated with a higher incidence of neurologic abnormalities. In this study we tried to determine the prognostic value of repeated CSF latex agglutination testing. Methods : We retrospectively evaluated 19 patients admitted to Ewha Womans University Mokdong Hospital for bacterial meningitis from January 1997 to June 2002. Bacterial meningitis was confirmed by a positive CSF culture and a positive CSF latex agglutination test. Repeated CSF examinations were done at three, seven, 14, 21 and 28 days after antibiotics therapy. Neuroradiologic studies were performed. Results : The mean age was $10.6{\pm}12.3months$(range; two to 33 months). The male to female ratio was 2.8 : 1. The causative organisms were Haemophilus influenzae type b 57.9%, Group B Streptococcus 21.1%, Streptococcus pneumoniae 15.7% and Escherichia coli 5.3%. Three days after the initiation of antibiotics therapy, repeated CSF latex agglutination tests persisted as positive in nine (47.4%) out of 19 cases, but all CSF cultures became negative. In those cases with negative latex agglutination tests three days after antibiotics therapy, neuroradiologic findings were completely normal. But, in cases with positive latex agglutination tests three days after antibiotics therapy, neuroradiologic abnormalities such as cerebral infarction, encephalomalasia occurred in 44.4%. Conclusion : Repeated CSF latex agglutination testing was valuable as a prognostic factor in bacterial meningitis. Neuroradiologic abnormalities may occur in cases with delayed clearance of CSF latex agglutination tests more often than in cases with negative latex agglutination tests three days after antibiotics therapy.

• Pediatric Infection and Vaccine
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• v.10 no.1
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• pp.81-86
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• 2003

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.

• Pediatric Infection and Vaccine
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• v.10 no.2
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• pp.159-166
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• 2003

Purpose : Streptococcus pneumoniae is part of the normal flora but is also responsible for causing many invasive diseases such as pneumonia, meningitis, and sepsis in addition to noninvasive diseases such as otitis in children. Multi-drug resistant strains has raised a lot of concern worldwide and thus the importance of prevention has been emphasized. We have analyzed the current serotypes and antibiotic sensitivity of each serotype as a baseline study to estimate the efficacy of the pneumococcal vaccine in Korean children. Methods : One hundred sixteen cases of pneumococcus cultured at Yonsei Medical Center from September 2001 to January 2003 were analyzed. The serotyping was done with the Quellung reaction and penicillin resistance was tested using the oxacillin disc diffusion method. Results : Pneumococcus were cultured from the sputum in 76 cases(65.5%), from the blood in 13 cases(11.2%), from the ear discharge in 12 cases(10.3%), from the throat in 7 cases(6.0%), from the nasal cavity in 2 cases(1.7%), and one case(0.9%) each from the cerebrospinal fluid, eye discharge, peritoneal fluid, post-operational wound, brain abscess, and catheter tip. Serotyping was possible with 98 cases and the following serotypes were found; 15 cases of type 19F(15.3%), 11 cases of 19A(11.2%), 8 cases of 11A(8.2%), 7 cases each of 6A, 14 and 3(7.1%), 6 cases each of 35, 6B and 23F(6.1%). Eighty two cases(70.7%) out of 116 cases were penicillin resistant and serotypes 19F, 19A, 11A, 23F, 6A, 9V constituted the majority, 48 cases(59.8%). These serotypes showed resistance to cotrimoxazole (74.4%), tetracycline(69.5%), and erythromycin(90.3%) as well. In the 22 cases cultured from children, 19A and 19F were found in 25.0%, 6A, 6B, and 23F in 10.0%, 11A, 14, 19, and 29 in 5.0%. Fifty percent(10/20) of the clinical isolates were represented in the current 7-valent pneumococcal protein conjugate vaccine, and 85%(17/20) when the cross-reacting serotypes were included. Penicillin resistance was found in 86.4%(19/22). Conclusion : The percentage of serotypes included in the 7 valent pneumococcal protein conjugate vaccine found in our study was 40.8% which was less than other prior studies. In anticipation of a change of pneumococcal serotypes, a nationwide multicenter study is needed before the initiation of pneumococcal vaccines in Korea.