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tot lol Flashcards

1
Q

in analiza ELISA, enzimele utilizate sunt:

A

POD (HRP)
PA (fosfataza alcalina)
GOD

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2
Q

ELISA:

substrat si produs pentru HRP:

A

OPD (orto phenilen diamina) - p portocaliu, 490 nm

TMB (tetrametil benzidina) - galben , 450 nm

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3
Q

ELISA:

substrat si produs pentru PA:

A

PNP (para nitrofenil fosfat) - galben, 405 nm

Metil umbeliferil fosfat - metil umbeliferona, 365 - 445 nm.

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4
Q

ce e PSA si metoda de obtinere

A

Prostate Specific Ag - cancer de prostata - metoda Sandwich

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5
Q

Hormonul sarcinii

A

hCG - gonadotropina corionica umana

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6
Q

NAD

A

nicotinamid adenin dinucleotid - coenzima dehidrogenari

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7
Q

PLP

A

piridoxal fosfat - coenzima (decarboxilari)

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8
Q

TPP

A

tiamin pirofosfat coenzima

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9
Q

FAD

A

flavin adenin dinucleotid - cofactor (merge cu GOD)

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10
Q

FMN

A

flavin mononucleotid - cofactor (cu AAO)

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11
Q

GOT, GPT

A

glutamat oxaloacetat transaminaza / transferaza (cu PLP)

glutamat piruvat transaminaza (cu PLP)

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12
Q

ALT - (GPT)
AST - (GOT)
sunt cu PLP

A

alanin aminotransferaza - glutamat piruvat aminotransferaza
aspartat aminotransferaza - glutamat oxaloacetat aminotransferaza
pridioxal fosfat

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13
Q

UI = unitatea de activitate enzimatica

A

cantitatea de E care catalizeaza transformarea unui micromol de S per minut in cond standard.

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14
Q

activitatea specifica

A

activitatea unui preparat enzimatic rap la 1 mg proteina (micromoli de S transf pe min per mg proteina) fie la 1 cm^3 extract sau de sol ce contine enzima

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15
Q

Kcat

A

nr de turnover = k2 exces de S

nr max de molec de S ce pot fi trans de o molec de E pe unit de timp

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16
Q

1 katal

A

mol/s = cantit de E care trans 1 mol de S per sec

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17
Q

Vo=

A

vo= Vomax * [S] / ( kM + [S] )

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18
Q

GCK

A

glucokinaza trans G6P in glicogen in ficat

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19
Q

Conditii analiza enzimatica

A 
saturare cu S, S = 10 kM sau 100 kM
exces co enzima - r de ord 0 in rap cu acestea
cond initiale
temp si pH optime
viteza de r dep doar de activ enz
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20
Q

Det ADH

A

etanol + NAD = acetaldehida + NADH

NADH absoarbe la 340 nm

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21
Q

ACP

ALP

A

fosfataza acida - prostata

fosfataza alcalina - ficat, oase - rahitism, icter

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22
Q

GMT / GGT

A

gama glutamil transferaza

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23
Q

E din sange

A

se prefera ser, special daca e electroforeza
accepta plasma heparinizata
unii anticoag (fluorura, EDTA, citrat) inhibitori necesita activatori (Ca2+)
nu pb hemolizate, lipemia
max 4 h de la recoltare, stocare la rece mai mult timp

24
Q

Det CK - creatin kinaza

A

creatin fosfat + ADP (inv CK) = creatina + ATP nu optic activi
ATP + glucoza (HK) = ADP + G6P analog
G6P + NADP (G6PDH) = 6 fosfogluconat + NADPH
NADPH la 340 nm

25
Det LDH
lactat dehidrogenaza L-lactat + NAD (LDH) = piruvat + NADH mai putine erori piruvat + NADH (LDH) = L-lactat + NAD mai speed 340 nm
26
Valori critice ALP
> ciroza, osteomalacie, rahitism | < tumori osoase, leziuni hepatice
27
Det ALP
1. fenolftalein suflat la pH = 10 2. fenil fosfat disodic HIDROLIZA da fenol la 520 nm rezultatul cu 4AAP 4 amino antipirina fenol cu gen fosfat = (ALP) = FENOL + FOSFAT fenol + 4 AAP = chestie oribila la 520 nm
28
Det ACP
hidroliza alfa naftil fosfat (ACP) = alfa naftol + fosfat alfa naftol cu cromogen = colorant azoic comp cromofori Fast Blue B, Fast red TR ac tartric inhiba PAP = det prin diferenta flueriometric: 4 metil umbeliferon fosfat (MUP)
29
Fast Blue B | Fast Red TR
``` blue = brentamina (o dianisidina diazotata) red = 4 cloro 2 metilbenzen diazoniu ```
30
AST (GOT)
transfera amino de la aspartat la alfa ceto acetat cu PLP aspartat + alfa ceto glutarat (AST) = oxaloacetat + glutamat oxaloacetat + NADH (MDH)= malat + NAD MDH = malat dehidrogenaza
31
Det ALT (GPT)
1 ala + alfa CG (ALT) = piruvat + glutamat piruvat + NADH (LDH) = lactat + NAD 2 ala + alfa CG (ALT) = piruvat + glutamat piruvat + O2 + PO4 (piruvat oxidaza) = acetil fosfat + CO2 + H2O2 H2O2 + fluorofor (o-fenilen diamina) (HRP) = comp fluorescent + H2O
32
Det GGT
gama glutamiltranferaza g glutamil de la peptide ca glutationul (GSH) catre alti AA g glutamil p nitroanilina +gly-gly (GGT) = p nitroanilina + g glutamil - gly-gly p nito anilina abs la 410 nm
33
Abs p nitroanilina
410 nm
34
Abs produs reacyie fenol si 4 AAP
520 nm
35
DET G6PDH
G6P + NADP () = 6 fosfogluconolactona + NADPH si reactie cu diaphorase resazurin + NADPH (diaphorase) = Resorufin + NADP astia sunt cromoforii
36
Det amilaza
1 cu Iod scade semnalul ca urm a cons amidon 2 p nitrofenil maltoheptaozida = p nitro fenol care in pH > 7,5 e galben anticorpii monoclonali inhiba AML salivara 3. AML legata de colorant - amyloze azure - hidroliza - compus ce abs la 595 nm
37
Absorbtie compus hidroliza amyloze azure
595 nm
38
BALB
dimercaptopropanol tertbutirat
39
DTNB , TNB , cine absoarbe la cat?
5,5', ditiobis 2 nitrobenzoic acid | 5 tio 2 nitrobenzoic acid - la 412 nm galben
40
Det lipaza
BALB (lipaza) = HS-C-C(SH)-C-OH + 3 CH3-CH2-CH2-COOH | R-SH + DTNB = R-S-TNB + TNB
41
Det ChE
Colinesteraza ATCh + apa (AChE) = TCh + CH3COOH TCh + DTNB = TCh-TNB + TNB (412 nm)
42
Det glucozei
1. Reducere in mediu alcalin a Cu2+ -> Cu2O ac fosfomolibdneic + Cu2O = albastru de molibden 2. o-toluidina + glucoza (ac acetic glacial) = baza Schiff la 630 nm 3. GOD + HRP 4. HK + G6PDH glucometre = cea cu GOD 2 H2O2 + fenol + 4 AAP (HRP) = chinon imina + 4 H2O rosie la 550 nm
43
metoda biuretului
CuSO4 KNaC4H4O6 mediu alcalin abs la 540 nm
44
metode det proteine
albumina: 1. precipitare cu saruri sau acizi 2. det cont triptofan indirecta ac glicoxilic + globuline = compus cromogen (540 nm) albumina = proteine totale - globuline 3. electroforeza (separare, colorare, det abs benzi, det procente fractii proteice) 4. met spectro cu coloranti anionici metilorange 5. din urina - bandolete cu albastru de bromfenol - se fac albastre (din galben) daca este pozitiv
45
det uree
cu monoxima da diazina si apa la 540 nm R Nessler - 450 nm NH3 + K[HgI4] + 3 OH- R Berthelot - 540 nm glutamat + NAD + H20 (glutamat dehidrogenaza) = NH4 + ALFA CG + NADH
46
Det creatinina
creatinina + ac picric = compus colorat (520 nm)
47
BUN / creatinina mare (mare / normal) mare (mare / mare) mic (mic / normal)
mare / normala = pre renala - insuf card, hipercatab mare / mare = post renal - obstructie urinara mic / normal = renal - dieta fara proteine, afectiuni hepatice
48
la cat abs ac uric
293 nm
49
Det ac uric
ac uric + O2 + apa (uricaza) = alantoina + CO2 + H2O2 | 293 nm
50
Det amoniac
glutamat + NAD + H20 (glutamat dehidrogenaza) = NH4 + ALFA CG + NADH
51
Det colesterol
1. hidroliza KOH alcoolic 2. extractia eter petrol 3. R cu ac sulfuric si anh acetica - verde enzimatic colesterol ester + apa (colesterol esteraza) = col + ac grasi col + O2 (col oxidaza) = 4 colesten 3 ona + H2O2 2 H2O2 + fenol + 4 AAP (HRP) = chinon imina + 4 H2O rosie 550 nm
52
La cat abs chinon imina
550 nm
53
Det tri (ne enzimatica pt ca enzimatica idk)
tri (KOH alcool) = glicerol + ac grasi glicerol + ac periodic = formaldehida formaldehida + ac cromotopic = coloratie albastra
54
Icter hemolitic
f mult urobilinogen, BU crescut, BC normal | scaun inchis, urina normala
55
Icter obstructiv
BU normal, BC mare, urina brun, urobilinogen scazut, scaun deschis, prezente saruri biliare si pigmenti biliari in urina
56
Icter hepatocelular
BU si BC mari, bilirub in urina, urobilinogen crescut, prezinta urme saruri biliare
57
Sensibilitate
variatia semnalului masurat corespunzator variatiei concentratiei analitului de masurat cu o unitate

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