m s h t m l shared microbiology service(m s h t m l共享微生物学服务).docVIP

m s h t m l shared microbiology service(m s h t m l共享微生物学服务).doc

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MYCOLOGY MANUAL TABLE OF CONTENTS LAB SAFETY 3 DAILY ROUTINE OF THE MYCOLOGY LAB 4 SPECIMEN COLLECTION AND TRANSPORTATION 5 PROCESSING OF SPECIMENS 5 ISOLATION AND IDENTIFICATION 6 I. Reading of cultures 6 II. Identification 6 A) FILAMENTOUS FUNGI 6 B) YEAST 11 C) NOCARDIA 13 REPORTING 14 STAINING METHODS 17 ISOLATOR 10 BLOOD CULTURE SYSTEM FOR DIMORPHIC FUNGI 18 APPENDIX I - CONVERSION (Converting Mycelial Phase of Dimorphic Mould to a Yeast Phase) 22 APPENDIX II - CORNMEAL TWEEN-80/OXGALL AGAR 23 APPENDIX III - DETERMINING CYCLOHEXIMIDE RESISTANCE OF AN ISOLATE 25 APPENDIX IV - FLUORESCENT MICROSCOPE (INSTRUCTIONS) 27 APPENDIX V - GERM TUBE TEST 28 APPENDIX VII - STOCK CULTURES -WATER CULTURE TECHNIQUE 30 APPENDIX VIII - API 20 CAUX - Yeast Identification System 31 APPENDIX IX - MEDIA / REAGENTS 33 Brain Heart Infusion Agar with 10% Sheep Blood, Gentamicin, Chloramphenicol, Cyclohexamide (BHIM) 33 BRAIN HEART INFUSION AGAR (BHIA) 34 CORNMEAL TWEEN 80 AGAR (OXOID) 35 ESCULIN BASE MEDIUM (EBM) pH 7.1 36 INHIBITORY MOLD AGAR (IMA) 38 LACTOPHENOL COTTON BLUE STAIN 40 MYCOSEL AGAR 41 OXGALL AGAR 42 POTATO DEXTROSE AGAR (PDA) 43 SODIUM PYRUVATE AGAR (NPA) FOR NOCARDIA 44 SABOURAUD AGAR MODIFIED (DIFCO) 45 SABOURAUD GENTAMICIN (50 mg/L) AGAR SLOPES 46 UREA AGAR SLOPES 47 10% POTASSIUM HYDROXIDE 48 APPENDIX X – Flow Charts for Identification and Reporting 49 Flow Chart 1 - Mycology Fungi Smear and Culture Reading 49 Flow Chart 2 - Direct Microscopy 49 Flow Chart 3 - Zygomycetes 49 Flow Chart 4 - Dematiaceous 49 Flow Chart 5 - Hyalohyphomycetes 49 Flow Chart 6 - White Mould 49 Flow Chart 7 - Aspergillus 49 Flow Chart 8 – Dimorphic Fungi 49 Flow Chart 9 - Actinomyces 49 Flow Chart 10 - Yeast 49 Flow Chart 11 - Dermatophytes 49 Record of Edited Revisions 50 LAB SAFETY Refer to Laboratory Safety Manual Note the following when processing Mycology isolates: ALL work on filamentous fungus is carried out in LAMINAR AIRFLOW BIOSAFETY CABINET TYPE 2. Bio safety Lev

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