1.Serotype Distribution and Antimicrobial Susceptibilities of Invasive Streptococcus pneumoniae Isolates from Adults in Korea from 1997 to 2012.
Chung Jong KIM ; Jin Su SONG ; Su Jin CHOI ; Kyoung Ho SONG ; Pyeong Gyun CHOE ; Wan Beom PARK ; Ji Hwan BANG ; Eu Suk KIM ; Sang Won PARK ; Hong Bin KIM ; Nam Joong KIM ; Eui Chong KIM ; Myoung Don OH
Journal of Korean Medical Science 2016;31(5):715-723
In Republic of Korea, a 7-valent pneumococcal conjugated vaccine (PCV7) was licensed for use in infants in 2003, and 13-valent PCV (PCV13) replaced it since 2010. We investigated trends in serotype distribution and antibiotic susceptibility of pneumococcal isolates from adult patients with invasive pneumococcal diseases (IPD). Invasive pneumococcal isolates from adult patients of ≥ 16 years of age were collected from 1997 to 2012. Serotypes of the isolates were determined by the Quellung reaction. Distribution of serotypes was analyzed according to the vaccine types. Antibiotic susceptibility was tested by using E-test strips. A total of 272 invasive pneumococcal isolates were included. The most common serotypes were serotype 19F (8.5%, 23/272), and serotype 3 (8.1%, 22/272), and 24.6% (67/272) of the isolates were of non-vaccine serotypes. Of the 272 isolates, 2.6% (7/272) were penicillin MICs of ≥ 4 µg/mL. The proportion of the PCV13 serotypes decreased from 63.3% (50/79) in 1997-2003 to 48.6% (17/35) in 2011-2012, whereas that of non-vaccine serotypes was 26.6% (21/79) and 25.7% (9/35), respectively, for the same periods. The proportion of the PCV13 serotypes showed a decreasing trend among adult patients with IPD over the study period.
Adolescent
;
Adult
;
Aged
;
Anti-Infective Agents/*pharmacology/therapeutic use
;
Ceftriaxone/pharmacology/therapeutic use
;
Female
;
Humans
;
Male
;
Microbial Sensitivity Tests
;
Middle Aged
;
Penicillins/pharmacology/therapeutic use
;
Pneumococcal Infections/drug therapy/*microbiology/mortality
;
Republic of Korea
;
Serogroup
;
Serotyping
;
Streptococcus pneumoniae/*drug effects/isolation & purification
;
Young Adult
2.Detection of the Efflux-Mediated Erythromycin Resistance Transposon in Streptococcus pneumoniae.
Azadeh AZADEGAN ; Ali AHMADI ; Abdolaziz Rastegar LARI ; Malihe TALEBI
Annals of Laboratory Medicine 2015;35(1):57-61
BACKGROUND: The present analysis focuses on phenotypic and genotypic characterizations of efflux-mediated erythromycin resistance in Streptococcus pneumoniae due to an increase in macrolide resistance in S. pneumoniae worldwide. METHODS: We investigated the prevalence of efflux-mediated erythromycin resistance and its relevant genetic elements from 186 specimens of S. pneumonia isolated from clinical and normal flora from Tehran, Iran. The presence of erythromycin resistance genes was tested by PCR with two sets of primers, specific for erm(B) and mef(A/E), and their genetic elements with tetM, xis, and int genes. Isolates were typed with the BOX PCR method and tested for resistance to six antibiotics. RESULTS: Antibiotic susceptibility tests revealed that 100% and 47% isolates were resistant to tetracycline and erythromycin, respectively. The erythromycin and clindamycin double-disc diffusion test for macrolide-lincosamide-streptograminB (MLSB) resistance phenotype showed 74 (84%) isolates with the constitutive MLSB phenotype and the remaining with the M phenotype. BOX PCR demonstrated the presence of 7 types in pneumococci with the M phenotype. Fourteen (16%) isolates with the M phenotype harbored mef(A/E), tetM, xis, and int genes. CONCLUSIONS: The present results suggest dissemination of polyclonal groups of S. pneumoniae with the M phenotype carrying resistance genes attributed to transposon 2009.
Anti-Bacterial Agents/*pharmacology
;
Bacterial Proteins/genetics
;
DNA, Bacterial/metabolism
;
Drug Resistance, Multiple, Bacterial/*genetics
;
Erythromycin/*pharmacology
;
Genotype
;
Humans
;
Microbial Sensitivity Tests
;
Phenotype
;
Pneumococcal Infections/microbiology/pathology
;
Polymerase Chain Reaction
;
Streptococcus pneumoniae/*drug effects/*genetics/isolation & purification
;
Tetracycline/pharmacology
3.Nasopharyngeal carriage rate, antimicrobial resistance and serotype distribution of Streptococcus pneumoniae among children with upper respiratory infection.
Sang-Jie YU ; Wei GAO ; Wei SHI ; Lin YUAN ; A-Dong SHEN ; Kai-Hu YAO ; Yong-Hong YANG
Chinese Journal of Contemporary Pediatrics 2014;16(10):988-992
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
4.Characterization of erythromycin-resistant Streptococcus pneumoniae isolates causing invasive diseases in Chinese children.
Xiang MA ; Kai-hu YAO ; Gui-lin XIE ; Yue-jie ZHENG ; Chuan-qing WANG ; Yun-xiao SHANG ; Hui-yun WANG ; Li-ya WAN ; Lan LIU ; Chang-chong LI ; Wei JI ; Xi-wei XU ; Ya-ting WANG ; Pei-ru XU ; Sang-jie YU ; Yong-hong YANG
Chinese Medical Journal 2013;126(8):1522-1527
BACKGROUNDErythromycin-resistant Streptococcus pneumoniae isolates that causing invasive pneumococcal diseases (IPD) in Chinese children remain uncharacterized. This study aims to identify the resistance genes associated with erythromycin resistance and to determine the genetic relationships of IPD isolates in Chinese children.
METHODSA total of 171 S. pneumoniae strains were isolated from 11 medical centers in China from 2006 to 2008. All the isolates were characterized via serotyping and antibiotic susceptibility determination. The erythromycin-resistant isolates were further characterized via ermB and mefA gene detection, multi-locus sequence typing analysis, and pulsed-field gel electrophoresis.
RESULTSA total of 164 (95.9%) isolates showed resistance to erythromycin, of which 162 strains with high high-level resistance (MIC ≥ 256 µg/ml). A total of 104 (63.4%) isolates carry the ermB gene alone, whereas 59 (36.0%) harbor both ermB and mefA genes. Of the 59 strains, 54 were of serotypes 19A and 19F and were identified as highly clonal and related to the Taiwan(19F)-14 clone.
CONCLUSIONSThe erythromycin resistance rate in IPD isolates is significantly high and is predominantly mediated by the ermB gene. Isolates that carry both ermB and mefA genes are predominantly of serotypes 19A and 19F.
Adolescent ; Anti-Bacterial Agents ; pharmacology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Erythromycin ; pharmacology ; Humans ; Infant ; Multilocus Sequence Typing ; Pneumococcal Infections ; microbiology ; Serotyping ; Streptococcus pneumoniae ; classification ; drug effects ; genetics ; isolation & purification
5.Changes in Serotype Distribution and Antibiotic Resistance of Nasopharyngeal Isolates of Streptococcus pneumoniae from Children in Korea, after Optional Use of the 7-Valent Conjugate Vaccine.
Eun Young CHO ; Hyun Mi KANG ; Jina LEE ; Jin Han KANG ; Eun Hwa CHOI ; Hoan Jong LEE
Journal of Korean Medical Science 2012;27(7):716-722
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
6.Clinical analysis of 20 cases with Streptococcus pneumoniae necrotizing pneumonia in China.
Jin-rong LIU ; Bao-ping XU ; Hui-min LI ; Ji-hang SUN ; Bao-lin TIAN ; Shun-ying ZHAO ; Zai-fang JIANG
Chinese Journal of Pediatrics 2012;50(6):431-434
OBJECTIVEStreptococcus pneumoniae necrotizing pneumonia (SPNP) was reported elsewhere but not in China yet. Inappropriate treatment due to poor recognition of this disease could influence its prognosis. This paper presents the clinical characteristics, diagnosis and treatment of SPNP hoping to elevate pediatrician's recognition level for this disease.
METHODClinical manifestations, radiological findings, treatment and prognosis of 20 patients (9 boys, 11 girls) who had been hospitalized with SPNP in Beijing Children's Hospital from 2004-2011 were retrospectively analyzed.
RESULTThe patients included in this study aged from 9 months to 6 years [(27.9 ± 15.8) m] and were healthy before admission. They were febrile for 8 to 50 days [(27.7 ± 13.5) d] and hospital day was 24 - 55 days [(36.5 ± 8.3) d]. The general condition of all subjects was relatively poor and they all had fever and cough. One child had moderate fever and nineteen children had high fever. Dyspnea was found in sixteen children. Fine rales were found on auscultation in 18 children, among whom diffuse wheeze appeared in 4 children, and wheezy phlegm was found in two children. Signs of pleural effusion were discovered in all cases by physical examination and chest X-ray. White blood cell (WBC) count was 16.2 - 60.95×10(9)/L and neutrophil was 70.5% - 80.2% in peripheral blood routine test. Erythrocyte sedimentation rate (ESR) was 44 - 109 mm/h [(69.6 ± 16) mm/h]and C-reactive protein (CRP) was 80 - > 160 mg/L. The pleural effusion biochemistry and routine test revealed a WBC count of 6400×10(6)/L-too much to count, polykaryocyte of 51% - 90%, glucose of 0.02 - 1.8 mmol/L, protein of 32 - 51 g/L and LDH of 5475 IU/L-or higher. Pleural effusion culture in all cases and blood culture in 2 cases was positive for Streptococcus pneumoniae. Chest X-ray or CT revealed high density and well-distributed lobar consolidation in one lung or two lungs initially. Single or multiple low density lesions in the area of lobar consolidation were found a week later, accompanied by multiple cystic shadow or cavity at the same time or afterwards. Bulla of lung appeared later. Pleural effusions were found in all patients. Seven cases complicated with hydropneumothorax, two with otitis media, one with heart failure, one with cardiac insufficiency. Seventeen patients were treated with vancomycin or teicoplanin or linezolid two with amoxicillin and clavulanate potassium. Other two patients had been treated with meropenem and cephalosporin antibiotics respectively before admission, and they had been at recovery stage when they were hospitalized. Thoracic close drainage and thoracoscopy were performed respectively in 18 cases and 3 cases, respectively. After a follow up of more than 6 months, chest CT showed that almost all lesions in lungs recovered during 4-6 months. No one received pneumonectomy.
CONCLUSIONSPNP has special manifestations. The incidence in infants is higher. Patients' general condition is poor and febrile course is relatively long. All patients manifested fever and cough, with a presence of dyspnea in most of them. WBC, neutrophil and CRP elevated apparently. The characteristic of pleural effusion indicates empyema. In early stage, the chest X-ray and CT showed high-density lobar lesions, followed by low-density lesions and cyst gradually. Bulla of lung and/or hydropneumothorax may appear at the late stage. But if diagnosed and treated promptly, the prognosis of SPNP was relatively good.
Anti-Bacterial Agents ; therapeutic use ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Dyspnea ; diagnosis ; drug therapy ; epidemiology ; Female ; Fever ; diagnosis ; drug therapy ; epidemiology ; Humans ; Infant ; Leukocyte Count ; Lung ; diagnostic imaging ; pathology ; Male ; Methylprednisolone ; therapeutic use ; Pleural Effusion ; diagnosis ; drug therapy ; epidemiology ; Pneumonia, Pneumococcal ; complications ; diagnosis ; drug therapy ; Prognosis ; Retrospective Studies ; Streptococcus pneumoniae ; drug effects ; isolation & purification ; pathogenicity ; Tomography, X-Ray Computed ; Treatment Outcome
7.Molecular epidemiology of serotype 19A Streptococcus pneumoniae isolated from children in Beijing, 1997-2006.
Lian XUE ; Kai-Hu YAO ; Sang-Jie YU ; Zun-Jie LIU ; Jing QIAN ; Xu-Zhuang SHEN ; Yong-Hong YANG
Chinese Medical Journal 2011;124(12):1769-1774
BACKGROUNDDespite the prevalence of Streptococcus pneumoniae serotype 19A, the molecular characteristics of this serotype are yet to be fully elucidated. The aim of this study was therefore to determine the homology of the serotype 19A in China.
METHODSPulsed-field gel electrophoresis and multilocus sequence typing were done to these forty-nine serotype 19A isolates to investigate the relationship between the strains prevalent in Beijing and other regions.
RESULTSFrom 1997 to 2006, the percentage of serotype 19A isolates increased. The susceptibility rate to penicillin and amoxicillin decreased and the resistance rate to cefuroxime increased. ST320 was the most prevalent ST, followed by ST3546. There were six new STs identified in our study. The serotype 19A strains were classified into six different pulsed-field gel electrophoresis (PFGE) patterns. ST320, which was associated with two different PFGE patterns (A and D), accounted for 32 isolates, and ST3546, which was associated with two PFGE patterns (B and E), accounted for eight isolates.
CONCLUSIONSFrom 2003 onwards, ST320 was the most common ST and the rate of resistance to cefuroxime increased significantly. Further long-term surveys of Streptococcus pneumoniae serotype 19A are required to monitor ST prevalence and antimicrobial resistance in this important human pathogen.
Child, Preschool ; China ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Infant ; Infant, Newborn ; Molecular Epidemiology ; Pneumococcal Infections ; drug therapy ; microbiology ; Serotyping ; Streptococcus pneumoniae ; classification ; drug effects ; genetics ; isolation & purification ; Time Factors
8.Pathogenic bacteria distribution and drug susceptibility in children with acute otitis media in Pearl River Delta.
Ruijin WEN ; Qiulian DENG ; Changzhi SUN ; Shengli GAO ; Jia TAO ; Renzhong LUO
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2011;25(19):884-887
OBJECTIVE:
To investigate the pathogenic bacteria distribution and drug susceptibility in children with acute otitis media (AOM) in different age and different season in the Pearl River Delta region.
METHOD:
Four hundred and forty-two children diagnosed as AOM were divided into three groups by age factor and four groups by season factor. Midge ear pus collecting and culturing were used for bacteria and antimicrobial susceptibility test.
RESULT:
(1) Strains of bacteria were isolated from 356 children with the positive rate of 80.5%. Streptococcus pneumoniae (39.2%), staphylococcus aureus (25.9%) and haemophilus influenzae (7.4%) were the most frequently isolated pathogens. (2) Streptococcus pneumoniae was the main pathogenic bacteria in 0-1 year group and > 1-3 years group (P < 0.05), staphylococcus aureus was the most frequently isolated pathogen in >3 years group (P < 0.05); (3) In the season groups, the number of children with AOM decreased significantly in July-September group. There was no significant difference of streptococcus pneumoniae distribution among the four groups (P > 0.05). Staphylococcus aureus was the main pathogen in January-March group (P < 0.05); (4) Drug sensitivity shown that linezolid and ofloxacin were most sensitive to streptococcus pneumoniae and staphylococcus aureus, and macrolides had a good therapy effect to haemophilus influenzae.
CONCLUSION
The pathogens distribution and drug susceptibility in children with AOM were varies in different age and different season. As a result, a treatment should be done based on the climate, environment, and pathogens distribution of a region.
Adolescent
;
Anti-Bacterial Agents
;
pharmacology
;
therapeutic use
;
Child
;
Child, Preschool
;
China
;
epidemiology
;
Female
;
Haemophilus influenzae
;
drug effects
;
isolation & purification
;
Humans
;
Infant
;
Male
;
Microbial Sensitivity Tests
;
Otitis Media
;
drug therapy
;
epidemiology
;
microbiology
;
Seasons
;
Staphylococcus aureus
;
drug effects
;
isolation & purification
;
Streptococcus pneumoniae
;
drug effects
;
isolation & purification
9.Evaluation of the BD Phoenix Automated Microbiology System SMIC/ID-2 Panel for Antimicrobial Susceptibility Testing of Streptococcus pneumoniae.
Kyo Kwan LEE ; Nam Hee RYOO ; Sung Tae KIM ; Seok Lae CHAE ; Hee Jin HUH
The Korean Journal of Laboratory Medicine 2009;29(3):212-217
BACKGROUND: With the emergence of antimicrobial resistance among Streptococcus pneumoniae, a more accurate and automated antimicrobial susceptibility testing method is essential. We evaluated the BD Phoenix Automated Microbiology System (Becton Dickinson Diagnostic Systems, USA) SMIC/ID-2 panel for antimicrobial susceptibility testing of S. pneumoniae. METHODS: A total of 113 clinical strains of S. pneumoniae (88 penicillin susceptible strains, 8 intermediate strains, and 17 resistant strains by 2008 CLSI criteria) were tested. Minimum inhibitory concentrations (MICs) for penicillin, cefotaxime, clindamycin, erythromycin, levofloxacin, trimethoprim/ sulfamethoxazole, tetracycline, and vancomycin were determined by Etest (AB Biodisk, Sweden) and Phoenix System. The results obtained by Phoenix system were compared to those obtained by Etest. RESULTS: The overall essential agreement of MICs (within one dilution of MICs) defined by the Phoenix and Etest was 92.3%. Neither very major errors nor major errors were produced, and minor errors were 6.5%. Minor errors were frequently observed in susceptibility testings for penicillin (22.1%), cefotaxime (12.4%), and trimethoprim/sulfamethoxazole (11.5%). CONCLUSIONS: The Phoenix SMIC/ID-2 panel provided a simple and rapid susceptibility testing for S. pneumoniae, and the results were in a good agreement with those of Etest. The Phoenix system appears to be an effective automated system in clinical microbiology laboratories.
Anti-Bacterial Agents/pharmacology
;
Bacterial Typing Techniques/instrumentation/methods
;
Drug Resistance, Bacterial
;
Microbial Sensitivity Tests/*methods
;
Reagent Kits, Diagnostic
;
Streptococcus pneumoniae/*drug effects/growth & development/isolation & purification
10.Surveillance of antimicrobial susceptibility of Streptococcus pneumoniae and Hemophilus influenzae isolated from children in Guangzhou area between 2003 and 2004.
Xu-qiang HUANG ; Zuo-yuan XIAO ; Li DENG ; Qiu-lian DENG ; Yong-qiang XIE ; Yong-hong YANG
Chinese Journal of Pediatrics 2006;44(6):441-444
OBJECTIVETo investigate the situation of antibiotic resistance of Streptococcus pneumoniae (Sp) and Hemophilus influenzae (Hi) clinical isolates from children in Guangzhou area.
METHODSThe authors cultured, isolated and identified the Sp and Hi strains from nasopharyngeal secretion of patients who visited Guangzhou Children's Hospital for upper respiratory tract infection between 2003 and 2004. K-B disc diffusion and E-test for antibiotic susceptibility were performed for these clinical isolates.
RESULTSTotally 172 and 484 strains of Sp and Hi were respectively isolated from nasopharyngeal secretions in the hospital. For Sp strains, the rates of resistance to penicillin, amoxicillin/clavulanic acid, ceftriaxone, cefuroxime, cefaclor, erythromycin, tetracycline, chloramphenicol, sulfamethoxazole/trimethoprim (SMZ/TMP), clindamycin and ofloxacin were 32.0%, 11.1%, 32.6%, 18.1%, 39.5%, 82.6%, 78.5%, 24.4%, 87.2%, 69.2% and 3.1%, respectively. The penicillin non-susceptible Sp (PNSSP) isolates showed higher rates of resistance to other antimicrobial agents such as other beta-lactam antimicrobial agents, erythromycin, and SMZ/TMP than those of penicillin susceptible Sp (PSSP) isolates. More than 90% of PNSSP were multidrug resistant strains. The average rate of beta-lactamase production among 484 strains of Hi was 29.5% (143/484). For Hi isolates, the rates of resistance to ampicillin, amoxicillin/clavulanic acid, ceftriaxone, cefuroxime, cefaclor, SMZ/TMP, tetracycline, chloramphenicol, azithromycin, and ofloxacin were 40.1%, 3.4%, 4.1%, 1.9%, 5.6%, 56.2%, 52.1%, 17.4%, 2.1%, and 0.6%, respectively.
CONCLUSIONThe antimicrobials resistant Sp and Hi isolated from children with respiratory tract infection in the area have become a severe problem. The rate of resistance to penicillin of Sp had been decreased compared with the last three years, but the rate of resistance to ceftriaxone of Sp increased, and the multidrug resistance rates of PNSSP was rather high. PNSSP was characterized by a multidrug-resistance to erythromycin, tetracycline and SMZ/TMP. beta-lactamase production and ampicillin resistance among the Hi isolates from children in the area had increased generally during the period 2003 - 2004. The Hi isolates were more susceptible to the second and the third generation cephalosporins, amoxicillin/clavulanic acid and azithromycin.
Anti-Bacterial Agents ; pharmacology ; Child, Preschool ; China ; Drug Resistance, Bacterial ; Female ; Haemophilus Infections ; drug therapy ; microbiology ; Haemophilus influenzae ; drug effects ; isolation & purification ; Humans ; Infant ; Infant, Newborn ; Male ; Microbial Sensitivity Tests ; Nasopharynx ; microbiology ; Pneumonia, Pneumococcal ; drug therapy ; microbiology ; Respiratory Tract Infections ; drug therapy ; microbiology ; Streptococcus pneumoniae ; drug effects ; isolation & purification

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