MIXING/

SUBSTITUTION
STUDIES
WEEK 13 HEMA LAB
INTERPRETATION

MIXING STUDIES - First step when evaluating unexpected prolong PT or

- Distinguish clotting time prolongation due to a
coagulation factor efficiency or an inhibitor
- Mix patient plasma and normal pooled plasma
and measure the clotting time that was initially
prolonged
- Tests performed on blood plasma to distinguish
factor deficiencies from factor inhibitors
- Ex. Lupus anticoagulant, or specific factor
inhibitors, such as antibodies directed against
factor VIII
PTT results is to rule out preanalytical interference,
presence of contaminating
- If the APTT or PT is corrected by hormonal plasma, a
factor deficiency is indicated
- If the APTT or PT is not corrected by the addition of
normal plasma immediately, a strong inhibitor is
indicated
- Weak or time dependent inhibitor is indicated by a
prolonged APTT or PT following incubation at 37˚ for
1 to 2 hours (Factor VIII inhibitor)

MIXING STUDY

- Patient plasma is mixed 1:1 with normal pooled

plasma
- Contains 100% of the normal factor level results in a
level of 50% in the mixture Plasma has fibrinogen; serum doesn’t have fibrinogen
- Correction with mixing indicates factor deficiency,
failure to correct an inhibitor
 FRESH AGED ADSORBED FRESH AGED
SUBSTITUTION/ PLASMA
12345
PLASMA
12347
PLASMA
1 3 4 5 8 11
SERUM
2 3 4 9 10
SERUM
3 4 9 10
7 8 9 10 9 10 11 12 13 11 12 11 12
MIXING STUDIES 11 12 13 12 13
58 2 7 9 10 1 5 8 13 1 2 5 8 13

WEEK 13 HEMA LAB FRESH PLASMA

- Coagulation factors present in this reagent: Factor I,

SECONDARY HEMOSTASIS
II, III, IV, V, VII, VIII, IX, X, XI, XII, XIII
- INTRINSIC FACTORS
AGED PLASMA
o 12,11,9,8
o PT= normal - Coagulation factors present in this reagent: Factor I,
o APTT= increased II, III, IV, VII, IX, X, XI, XII, XIII
- EXTRINSIC FACTORS - Absent: Factor V, VIII [heat-labile]
o TF,7
o PT= increase ADSORBED PLASMA
o APTT= normal
- COMMON FACTORS - Coagulation factors present in this reagent: Factor I,
o 10,5,2,1 III, IV, V, VIII, XI, XII, XIII
o PT and APTT= increase - Absent: Factor II, VII, IX, X

FIBRINOGEN PROTHROMBIN CONTACT GROUP FRESH SERUM

GROUP GROUP
Factor I: Factor II: Factor XI: Plasma- - Coagulation factors present in this reagent: Factor II,
Fibrinogen Prothrombin Thromboplastin III, IV, IX, X, XI, XII
Antecedent - Absent: Factor I, V, VIII, XIII
Factor V: Labile Factor VII: Stable Factor XII:
factor factor Hageman factor AGED SERUM
Factor VIII: anti- Factor IX: Pre-K
hemophilic factor Christmas factor - Coagulation factors present in this reagent: Factor III,
Factor XIII: fibrin Factor X: Stuart- HMWK IV, IX, X, XI, XII
stabilizing factor Prower Factor - Absent: Factor I, II, V, VIII, XIII

Highlight- nonabsorbable factors; not present in absorbed

plasma

Highlight- Absent in fresh serum

Prothrombin: Factor II: as serum ages nawawala

Not corrected: prolonged test; deficiency in absent

coagulation proteins

Corrected: deficiency in coagulation proteins present in the

reagent

only PT abnormal or prolonged- no longer perform mixed

study= factor 7 lang problem

Tissue factor- hindi nagiging deficient; lahat ng tao meron

PROLONGED APTT
PROLONGED PT

Reagent pool plasma

Ex. Prolonged or abnormal= perform mixing study= para

malaman bakit prolonged

NPP: 1:1

Measure immediately or incubate for 1-2 hours at 37˚C

MIXING STUDY
Mix dalawa:
- Clot-based test in which 1 part normal plasma and 1
part patient plasma is combined then tested by the Corrected- nagging normal na from being
partial thromboplastin time (PTT), immediately or
after an incubation period - Added reagent; present yung deficient kay patient

Fail to correct

- Absent yung deficient

MIXING STUDY

- A factor deficiency of the patient will show a

correction to a normal PPT for the mix
 - An inhibitor in the patient plasma will not correct to a
normal PTT with the mix

SUBSTITUITION/ MIXING STUDIES

- Clot-based test
- Used to assess the abnormal or prolonged results of
clotting time tests
- Performed on citrated plasma (PPP) to determine
deficiencies of coagulation factors

9, 10,7,2- di agad naabsorb

Fibrinogen- hindi present in serum

Serum- liquid part of clotted blood

Plasma- liquid part of uncoagulated- with Fibrinogen

Reagent: normal reagent plasma

Fresh plasma
Aged plasma

Adsorbed plasma

Fresh serum

Aged serum

Corrected- present sa AP

Gray shaded- coagulation factors present

Light pink shaded- absent coagulation factors

5 and 8- heat labile

Bakit daw absent yung sa adsorbed aged serum

NC- absent nasa 5 and 8 ng AP

4- not included kasi calcium

FS:

C- 2,3,9,10,11,12

NC- 1,5,8,13

IN- 12,11,9,8

FP- 1,2,3,5,8,7,9,10,11,12,13

AP- 1,3,5,8,11,12,13

Lahat ng present kay aged serum icacancel natin

3,9,10,11,12- icancel natin

!!!!!!

IN- 12,11,9,8

AP=C: 1,3,5,8,11,12,13

AS= NC: 1,2,5,8,13

Common denominator:8= factor 8 deficient

Fibrinogen and 5 and 10- common factor

=8

aPTT- plasma; label

PT abnormal only; no longer perform mixed stufy= factor 7

lang

IN: 12,11,9,8

AP: 2,7,9,10

Longer method AS: 3,9,10,11,12

9 deficient
Common: 10,5,2

AP: 2,7,9,10

AS: 1,2,5,8,13

Factor II

Hindi lagging ptt lang minemeasure

Usually Ptt- mas madamin in factors involve

PT- factor 7 alone

Hindi nacoorect- may inhibitor

Correct?=