PCR法による肺炎球菌の薬剤耐性迅速スクリーニング法の検討

PCR法检测肺炎球菌耐药性快速筛查方法

• 批准号: 08770611
• 负责人: 長井 健祐
• 金额: $ 0.51万
• 依托单位: Kurume University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-08770611/
• 关键词: 肺炎球菌; 薬剤耐性; PCR; 耐性スクリーニング法

1993年分離の肺炎球菌98株は,寒天平板希釈法(標準法)では高度耐性(PCG;MIC≧2.0μg/ml)10株,中等度耐性(0.13-1.0μg/ml)35株,感性株(MIC≦0.06μg/ml)53株であった.この結果を以下の3つの方法の結果と比較した.1 Oxacillin(MPIPC)disk:Penicillin耐性株45株は,すべてMPIPC diskで耐性と判定された.感性株53株中,MPIPC diskで耐性と判定された株が12株にみられ,標準法との一致率は87.8%(95%CI:79.6-93.5%)であった.2 E test:高度耐性株10株のうち,E testで5株が中等度耐性と判定された.中等度耐性株35株では,E testで7株が高度,1株が感性と判定された.感性株53株では,E testで7株が中等度耐性と判定された.標準法との一致率は79.6%(95%CI:70.3-87.1%)であった.3 PCR法による薬剤耐性遺伝子の検出:Penicllin高度耐性株10株中,7株でPBP2B class B変異,PBP2X変異遺伝子の両方が検出された.残りは2株でclassB変異のみ検出され,1株でPBP2X変異とPBP2B感性型遺伝子(class S)が検出された.中等度耐性株35株中,30株でPBP2B classB変異,PBP2X変異のいずれかが検出され,class S遺伝子のみ検出された株が5株であった.耐性株だけの薬剤感受性と耐性遺伝子の一致率は,88.9%(95%CI:76.0-96.3%)であった.髄膜炎などの重症感染症分離の肺炎球菌では,まずMPIPC diskとE testを行い判定に最低18時間かかるため,数時間で結果が判明するPCR法で耐性遺伝子の確認を合わせて行う事が望ましいと考えられた.

98 strains of pneumococci isolated in 1993 were highly resistant (PCG;MIC ≥ 2.0μg/ml)10 strains, moderately resistant (0.13-1.0μg/ml)35 strains, susceptible (MIC ≤ 0.06μg/ml)53 strains. The results of the following 3 methods were compared. 1 Oxacillin(MPIPC)disk:Penicillin resistant strain 45, MPIPC disk resistant strain determination. Among the 53 susceptible plants,MPIPC disk was used to determine the tolerance of 12 plants. The agreement rate of standard method was 87.8%(95% CI:79.6-93.5%). E test: highly tolerant plants 10 plants were used to determine the tolerance of 5 plants. 35 plants with medium tolerance, 7 plants with high resistance and 1 plant with sensitive resistance. 53 sensitive plants,E test, 7 moderate tolerant plants. The concordance rate of standard method was 79.6%(95% CI:70.3-87.1%). 3 PCR method was used to detect the resistant strains: 7 strains of Peniclin highly resistant strains had PBP2B class B variant, and the PBP2X variant strains were detected. Two classB strains were detected, and one PBP2X strain was detected. Among the 35 plants with medium tolerance, 30 plants showed PBP2B class B variant, PBP2X variant and 5 plants showed S class variant. The concordance rate between susceptibility and tolerance was 88.9%(95% CI:76.0-96.3%). Pneumococcus isolated from severe infections due to meningitis was identified by MPIPC disk and E test at least 18 times. PCR method was used to confirm the resistance vector.