Isolates were classified into 3 age groups: group 1: children <5 years with isolates from both sterile sites (total 64: 59 blood, 4 cerebrospinal fluid, 1 pleural fluid) and non-sterile sites (total 42: 32 respiratory specimen, 6 ear swab, 2 eye swab, 2 gastric wash), group 2: patients 5–64 years with isolates from sterile sites only (total 62: 53 blood, 3 cerebrospinal fluid, 6 pleural fluid), and group 3: patients >65 years with isolates from sterile sites only (total 46: 44 blood, 2 pleural fluid). In this study, we performed serotyping and analysed serotype this website coverage of PCV-7, PCV-9, PCV-10, PCV-11 and PCV-13. PCV-9 is PCV-7 plus 1 and 5. PCV-10 is PCV-9 plus 7F, PCV-11 is PCV-10
plus 3, PCV-13 is PCV-11 plus 6 A and 19A. To determine capsule serotypes of isolates, we performed the Quellung test , using various specific group and factor antisera according to the manufacturer’s guideline from the State Serum Institute, Denmark. Typing was done with an addition of a loopful (a few microliters) of methylene blue 0.3% (w/v) in a bacterial suspension on a glass slide, using a microscope (OYMPUS BX 50 Model U-MD08, Oympus Corporation, Tokyo, SB203580 nmr Japan) with an oil immersion
lens (magnification, 10 × 100). The isolates that were not one of the serotypes included in PCV-7, PCV-9, PCV-10, PCV-11 and PCV-13 vaccines were not further typed and were labeled as nonvaccine types. Bacterial susceptibility of the isolates to penicillin, cefotaxime, ofloxacin and ciprofloxacin were evaluated by standard microbroth dilution using cation-adjusted Mueller-Hinton broth supplemented with 3% lysed horse blood  and E-test method (AB Biodisk, Sweden) according to the manufacturer’s guideline. S. pneumoniae ATCC 49619 was used as the control. The penicillin minimal inhibitory concentrations (MIC) were interpreted as susceptible, intermediate or resistant category according to Clinical Laboratory Standards Institute (CLSI) recommendations . This new criteria take into account whether penicillin is given orally or parenterally and whether a patient has meningitis.
Under the former criteria, the isolates from all clinical syndrome and penicillin routes, were interpreted as susceptible, intermediate, and resistant if MIC were ≥0.06, 0.12–1, and ≥2 μg/ml, respectively. Under the new criteria, the isolates are classified into 3 categories, almost i.e., meningitis with parenteral penicillin treatment (susceptible and resistant if MIC are ≤0.06 and ≥0.12 μg/ml, respectively); nonmeningitis with parenteral penicillin treatment (susceptible, intermediate and resistant if MIC are ≤2, 4 and ≥8 μg/ml, respectively); and non-meningitis with oral penicillin treatment (susceptible, intermediate, and resistant if MIC were ≤0.06, 0.12–1, and ≥2 μg/ml), respectively. The criterion for resistance to ciprofloxacin was MIC ≥4 μg/ml ; S. aureus ATCC 25923 was used as the control. The descriptive analysis was used in this study.