腸內桿菌鑑定
(EMB, MacConkey, Ureas)
抗酸性染⾊
嗜⾎桿菌
江倪全
https://sites.google.com/a/gap.cgu.edu.tw/bac_pathog/lecture
2018-04-24
學⼠後護理系
細菌⽣化鑑定的概念
尤加利樹
選擇性培養基、鑑別性培養基、滋養培養基
✓
選擇性培養基
(Selective media)
-
選擇性培養特定族群細菌
(e.g. G+ or G-)
✓
鑑別性培養基
(Differential media)
-
加以區分特定族群中之細菌種類
✓
滋養培養基
(Enriched media)
MacConkey agar plate
✓
培養基成份與功用
-
結晶紫
(Crystal violet)
-
膽
(Bile salts)
✓
抑制革蘭氏陽性菌生長
-
乳糖
(Lactose)
✓
乳糖發酵菌:
-
E. coli
✓
非乳糖發酵菌
-
沙門氏菌
(
Salmonella
)
-
志賀氏菌
(Shigella
)
-
Neutral red
✓
酸 指示劑 (acid: red)
Escherichia coli and Salmonella typhimurium
Eosin-methylene blue (EMB) agar plate
伊紅 基藍培養基
✓
培養基成份與功用
-
基藍
(Methylene blue)
✓
抑制革蘭氏陽性菌生長
-
伊紅
(Eosin)
✓
抑制革蘭氏陽性菌生長
✓
酸 指示劑: 中性至酸性—
無色至黑色
-
乳糖
(lactose)
；蔗糖
(Sucrose)
Escherichia coli
Escherichia coli
IMViC test: Indole, Methyl red, Voges-Porskauer,
Citrate test
✓
Indole test:
吲哚
產生試驗
✓
Methyl red (MR) test:
葡萄糖發酵試驗
✓
Vogest-Proskauer (VP) test:
弗普二氏試驗
Indole test:
吲哚產⽣試驗
Negative Positive Uninoculated
Tryptophan Indole + Pyruvic acid + Ammonia
Tryptophanase
Kovac’s reagent
+ Quinoidal red-violet compound
p-dimethylaminobenzaldehyde Indole
HCl
Isoamyl alcohol
MR-VP test
Negative Positive
Uninoculated
Glucose
+
H
2O
Lactic acid
Acetic acid
Formic acid
CO
2H
2(pH 4)
+
+
Methyl red (MR) test
Glucose O2 Acetic acid 2,3-butanediol
acetylmethylcarbinol CO2 H2 (pH 6.0)
+ + +
Barritt’s reagent
Acetylmethylcarbinol α-naphthol
40% KOH
Diacetyl Guanidine (Pink complex)
+ +
(e.g. E. coli)
(e.g. Enterobacter aerogenes)
Citrate test:
檸檬酸利用試驗
Negative Positive
Citrate
Citrase
Oxaloacetic acid + Pyruvic acid + Acetic acid + CO2
CO2 2Na+ H2O Na2CO3
Alkaline pH
+ +
尿素酶產⽣試驗
(Urease test)
Urea
+2H
2O
CO
2 +H
2O
+2NH
3Urease
快速微生物檢驗套組
API (Analytical profile index) system
快速微生物檢驗套組
抗酸性染色
(Acid fast staining)
G (+)
G (—)
Plasma membrane Peptidoglycan Arabinogalactan
Mannose-capped lipoarabinomannan
Mycolic acids Glycolipid
抗酸性染色
(Acid fast staining)
✓
抗酸性染色陽性菌
-
分枝桿菌屬
-
奴卡氏菌屬
✓
初染劑
-
石碳酸複紅
(Carbolfuchsin)
✓
脫色劑
-
酸性酒精
(3% HCl + 95% Ethanol)
✓
複染劑
-
亞 藍 (藍色)
(Methylene blue)
http://classes.midlandstech.edu
嗜⾎桿菌
(
Haemophilus
influenzae
)
✓
嗜⾎桿菌：營養苛求菌
-
⽣⾧所需養分
：
Hemin (X factor)
和
Nicotinamide adenine
dinucleotide (NAD, V factor)
-
X factor and V factor
：
紅⾎球內
巧克⼒培養基
chocolate agar
綿⽺⾎培養基
