实验八 细菌的鉴定(BBL半自 动鉴定系统) 一、教学要求 了解不同的微生物快速鉴定方法及应用 初步掌握细菌自动鉴定系统 (BBL CRYSTAL)的操作 技术 二、实验原理 1、 Miniaturized Biochemical Assays Bacteria can be identified by specific biochemical products they produce during growth. Biochemical and enzymatic reactions Miniaturized biochemical assays test for these specific products
实验八 细菌的鉴定(BBL半自 动鉴定系统) 一、教学要求 了解不同的微生物快速鉴定方法及应用 初步掌握细菌自动鉴定系统 (BBL CRYSTAL)的操作 技术 二、实验原理 1、 Miniaturized Biochemical Assays Bacteria can be identified by specific biochemical products they produce during growth. Biochemical and enzymatic reactions Miniaturized biochemical assays test for these specific products
API- Identification Kits Gram-negative bacteria Staphylococci
API- Identification Kits Gram-negative bacteria Staphylococci
2、Immunoassays ⚫ 血清学反应(Serological reactions) Agglutination reactions、Precipitation reactions ⚫ 免疫标记(immunolabel) Radioimmunoassays (RIA) Fluorescent immunoassays (FIA) Enzyme-linked immunosorbent assays (ELISA) Chemiluminesent Immunoassay(CLIA)、 Time-resolved fluorescent immunoassays(TRFIA) Immunogold labelling
2、Immunoassays ⚫ 血清学反应(Serological reactions) Agglutination reactions、Precipitation reactions ⚫ 免疫标记(immunolabel) Radioimmunoassays (RIA) Fluorescent immunoassays (FIA) Enzyme-linked immunosorbent assays (ELISA) Chemiluminesent Immunoassay(CLIA)、 Time-resolved fluorescent immunoassays(TRFIA) Immunogold labelling
ELISA(enzyme linked immunosorbent assay) The ELISA can be used to detect the presence of antigens that are recognized by an antibody, or it can be used to test for antibodies that recognize an antigen. ⚫ Sandwich ELISA
ELISA(enzyme linked immunosorbent assay) The ELISA can be used to detect the presence of antigens that are recognized by an antibody, or it can be used to test for antibodies that recognize an antigen. ⚫ Sandwich ELISA
Immunogold labelling techiques ⚫ 斑点免疫金渗滤法 ⚫ 胶体金标记免疫层析法
Immunogold labelling techiques ⚫ 斑点免疫金渗滤法 ⚫ 胶体金标记免疫层析法
Gold Tust Control 点orbent A起orbent 正ed band band Positif Goli Test Control Adsorbent 点orbent Negatif Pad band band Goll Test Control 点sorben Adsorbent Pad band band
3、Nucleic acid based assay ⚫ DNA hybridization – followed by detection with ⚫ Dot blot assay ⚫ Fluorescence or Chemiluminesence ⚫ Polymerase Chain Reaction (PCR) – followed by detection with ⚫ Gel electrophoresis ⚫ Fluorescence or Chemiluminesence ⚫ New assay formats under development
3、Nucleic acid based assay ⚫ DNA hybridization – followed by detection with ⚫ Dot blot assay ⚫ Fluorescence or Chemiluminesence ⚫ Polymerase Chain Reaction (PCR) – followed by detection with ⚫ Gel electrophoresis ⚫ Fluorescence or Chemiluminesence ⚫ New assay formats under development
Automated Identification Systems •VITEK系列全自动微生物分析系统 (biosMerieumx) •BBL CrystalTM半自动/自动细菌鉴 定系统(BD) 鉴定板,比浊仪,微生物鉴定专家数据分析系 统,自动判读仪(Autoreader)
Automated Identification Systems •VITEK系列全自动微生物分析系统 (biosMerieumx) •BBL CrystalTM半自动/自动细菌鉴 定系统(BD) 鉴定板,比浊仪,微生物鉴定专家数据分析系 统,自动判读仪(Autoreader)
1、 鉴定版(6类) NH 奈瑟氏/嗜血杆菌 3.0麦氏浊度 35~37℃ 孵育4小时 GP 革兰氏阳性菌 0.5麦氏浊度 35~37℃ 孵育18~24小时 RGP 快速革兰氏阳性菌 2.0麦氏浊度 35~ 37℃ 孵育4小时 ANR 厌氧菌 4.0-5.0麦氏浊度 35~37℃ 孵育4小时 ENF 肠/非发酵菌 0.5麦氏浊度 35~37℃ 孵育18~20小时 RSE 快速粪/肠菌 0.5-1.0麦氏浊度 35~37℃ 孵育3小时
1、 鉴定版(6类) NH 奈瑟氏/嗜血杆菌 3.0麦氏浊度 35~37℃ 孵育4小时 GP 革兰氏阳性菌 0.5麦氏浊度 35~37℃ 孵育18~24小时 RGP 快速革兰氏阳性菌 2.0麦氏浊度 35~ 37℃ 孵育4小时 ANR 厌氧菌 4.0-5.0麦氏浊度 35~37℃ 孵育4小时 ENF 肠/非发酵菌 0.5麦氏浊度 35~37℃ 孵育18~20小时 RSE 快速粪/肠菌 0.5-1.0麦氏浊度 35~37℃ 孵育3小时
三 材料与器材 ⚫ 菌种:待测的分纯菌株 ⚫ 试剂:无菌水,液体石腊,灭菌滴管等 ⚫ API 20 E 系统
三 材料与器材 ⚫ 菌种:待测的分纯菌株 ⚫ 试剂:无菌水,液体石腊,灭菌滴管等 ⚫ API 20 E 系统