Experiment IV Microbiological examination of bacteria Pan Tingting, PhD Key lab of Medical Molecular virology, Shanghai Medical College, Fudan University
Experiment Microbiological examination of bacteria IV Pan Tingting, PhD Key Lab of Medical Molecular Virology, Shanghai Medical College, Fudan University
Contents 1. Video Lab diagnosis of bacterial infection 2. Discussion Diagnostic principle of bacterial infection b) How to write a recorder of an experiment 3. Operation a) isolate bacterium from stool(lst): inoculation onto Mac con key agar b) Acid-fast stain 4. Design of widal test 5. Demonstration Mycobacterium and its colonies
Contents 1. Video Lab diagnosis of bacterial infection 2. Discussion a) Diagnostic principle of bacterial infection b) How to write a recorder of an experiment 3. Operation a) isolate bacterium from stool(1 st ): inoculation onto MacConkey agar b) Acid-fast stain 4. Design of widal test 5. Demonstration Mycobacterium and its colonies ： ： ： ：
Diagnosis of bacterial Infection Patient Clinical Non-microbiological diagnosis investigations Radiology Haematology Biochemistry Sample Take the correct specimen Take the specimen correctly Label package the specimen up correctly Appropriate transport storage of specimen A proper clinical assessment is essential for optimal use of laboratory services!
Diagnosis of Bacterial Infection Patient Clinical diagnosis Non-microbiological investigations Sample Take the correct specimen Take the specimen correctly Label & package the specimen up correctly Appropriate transport & storage of specimen A proper clinical assessment is essential for optimal use of laboratory services! Haematology Biochemistry Radiology
Principle of collection asepsIs avoid contamination by normal flora or bacteria from environment 2. take the correct specimen specimens include blood, urine, pus, stool, sputum, CSF, or other material, as indicated by the localization of the disease process 3. early before using antibiotics 4. fresh test ASAP (As Soon as possible
Principle of collection 1． 2. 3． ． asepsis avoid contamination by normal flora or bacteria from environment. take the correct specimen specimens include blood, urine, pus, stool, sputum, CSF, or other material, as indicated by the localization of the disease process. early before using antibiotics 4 fresh test ASAP（As Soon As Possible ）
Disease or symptoms specimens septicemia Blood specimen espiratory tract infection Sputum specimen urinary infection mid-stream urine digestive tract infection vomitus or stool purulent infection us secreta form STD Urogenital Tract epidemic meningitis CSF or blood enteric fever Blood(1-2weeks) Stool (2-3weeks
septicemia epidemic meningitis enteric fever Blood specimen Disease or symptoms specimens respiratory tract infection Sputum specimen urinary infection mid-stream urine digestive tract infection vomitus or stool purulent infection pus STD secreta form Urogenital Tract CSF or blood Blood (1-2weeks) Stool (2-3weeks)
Positive rate of specimens from patients suttering enteric fever 90 Agglutinins 80 blood 70 stool 60 urine 50 40 30 20 10 4 weeks post typhoid fever
0 10 20 30 40 50 60 70 80 90 1 2 3 4 5 6 7 8 weeks post typhoid fever positive rate(%) blood stool urine Positive rate of specimens from patients suffering enteric fever Agglutinins
Common pathogenic bacteria in clinical specimens specimen Common pathogenic bacteria blood Bacteroides fragilis(脆弱类杆菌) Brucella(布鲁氏菌) Burkholderia pseudomallei(类鼻疽伯克氏菌) Candida albicans and Cryptococcus neoformans(白色念珠菌和新型隐球菌 Haemophilus influenzae(流感嗜血杆菌) Neisseria meningitidis(脑膜炎奈瑟球菌 Non- ermenters other than Pseudomonas aeruginosa(铜绿假单胞菌) Other Enterobacteriaceae(肠杆菌) Pseudomonas aeruginosa Salmonella ty phi and non-ty phi(沙门氏菌) Staphy lococcus aureus(金黄色葡萄球菌) Streptococci(S. pyogenes, S. pneumoniae, viridans streptococci)(链球菌) Aeromonas and Plesiomonas, campylobacter spp, salmonella typhi, shigella (产气单胞菌、邻单胞菌、弯曲杆菌、伤寒沙门氏菌、志贺氏菌….) CSF Cryptococcus neoformans(新型隐球菌) Enterobacteriaceae(肠杆菌) Haemophilus influenzae(流感嗜血杆菌) Listeria monocytogenes(单核细胞增多性李斯特氏菌)
specimen Common pathogenic bacteria blood stool CSF Bacteroides fragilis Brucella Burkholderia pseudomallei Candida albicans and Cryptococcus neoformans Haemophilus influenzae Neisseria meningitidis Non-fermenters other than Pseudomonas aeruginosa Other Enterobacteriaceae Pseudomonas aeruginosa Salmonella typhi and non-typhi Staphylococcus aureus Streptococci (S. pyogenes, S. pneumoniae, viridans streptococci) （脆弱类杆菌） （布鲁氏菌 ） （类鼻疽伯克氏菌） （白色念珠菌和新型隐球菌 ） （流感嗜血杆菌 ） （脑膜炎奈瑟球菌） （铜绿假单胞菌 ） （肠杆菌 ） （沙门氏菌 ） （金黄色葡萄球菌） （链球菌） Aeromonas and Plesiomonas, campylobacter spp, salmonella typhi, shigella… … Cryptococcus neoformans Enterobacteriaceae Haemophilus influenzae Listeria monocytogenes （产气单胞菌 、邻单胞菌、弯曲杆菌、伤寒沙门氏菌、志贺氏菌 ） （新型隐球菌） （肠杆菌 ） （流感嗜血杆菌） （单核细胞增多性李斯特氏菌） Common pathogenic bacteria in clinical specimens
sputum throat swab urine Genital secretion wound stomach
sputum throat swab urine Genital secretion wound stomach
Lab diagnosis of bacterial infection bacteria tranditional method--isolate and identify the bacteria rapid diagnosis-—-1、 morphology 2, antigen test (protein) 3 nuclear acid(DNA/RNA antibodies Test the antibodies in patients using the known antigens
Lab diagnosis of bacterial infection bacteria tranditional method——isolate and identify the bacteria rapid diagnosis——1 morphology 2 antigen test (protein) 3 nuclear acid (DNA/RNA) antibodies Test the antibodies in patients using the known antigens ： ： 、 、
Serological test Test the antibodies in patients using the known antigens a antigen structure of bacter ia(o, h, Vi) toxin(o streptolys in) Specific anti gen a ant ibody early igM, later IgG
Serological test Test the antibodies in patients using the known antigens q n n q antigen：structure of bacteria（O,H,Vi） toxin（O streptolysin） Specific antigen antibody：early IgM，later IgG