CLINICAL CHEMISTRY POST-LABORATORY

DISCUSSION

LACTATE
DEHYDROGENAS
E
Nicole Sabado & Ma. Leila Galaura
 OBJECTIVES
At the end of the activity, the students must be able to:
Understand and apply the principles and concepts of
lactate dehydrogenase analysis.
Understand how to perform Lactate Dehydrogenase analysis
Understand the concepts presented in the review questions
associated with lactate dehydrogenase
WHAT IS THE E.C.
NUMBER FOR
LACTATE
DEHYDROGENASE?
A.
1.1.1.27

B. 3.2.1.1;
 LACTATE
E.C.
DEHYDROGENASE1.1.1.27
This is a hydrogen transfer enzyme that catalyzes the
oxidation of l-lactate to pyruvate with the mediation of
NAD+ as a hydrogen acceptor. It has a molecular weight of
134,000 Da. Two different polypeptide chains, designated H
(heart) and M (muscle), combine in five arrangements to
yield the five major isoenzyme fractions. These isoenzyme
fractions include LDH-1, LDH-2, LDH-3, LDH-4, and LDH-5.
A sixth LDH isoenzyme has been identified, which migrates
cathodic to LDH-5. LDH-6 is alcohol dehydrogenase.
 WHAT ARE THE TISSUE SOURCES FOR LACTATE
DEHYDROGENASE?

SKELETAL
HEART LIVE MUSCLE
ERYTHROCYTES

SMOOTH
LUNGS BRAI
MUSCLE N
 Principle of the
LDH catalyzes the interconversion of
lactic Test andpyruvic acids using the coenzyme
NAD.

Pyruvate + NADH + H+ --> Lactate + NAD+

Forward method: Wacker (pH 8.8)

NAD+ ---> Lactate Pyruvate

Reverse Method: Wrobleuski La Due (pH 7.2)

Pyruvate ----> Lactate
 MATERIALS AND
TEST
PROCEDURE
Patient’s serum
Lactate
Stopwatch
Test tubes
Dehydrogenase Micropipetto
reagent kit r Incubator
Spectrophotomete
r

..\Downloads\Lactate Dehydrogenase Test LDH Tes

t Procedure.mp4

..\Downloads\Lactate Dehydrogenase Assay Kit.mp

4
 REVIEW
QUEST ION
S
 WHAT ARE THE LD ISOENZYMES?
DIFFERENTIATE ONE FROM THE OTHER.
WHAT ARE THE LD ISOENZYMES?
DIFFERENTIATE ONE FROM THE
OTHER.
 Explain what is flipped
ratio.
Normally LD2 is greater than LD1 for a healthy
patient. Flipped Ratio occurs when the amount of
LDH-1 surpasses the amount of LDH-2 in the
serum. This happens in conditions involving cardiac
necrosis (AMI) and intravascular hemolysis. If ever
the patient has just experienced Myocardial
Infraction within 24 hours, the LD1 increases
resulting to a flipped ratio. The ratio should be less
than 1 but if MI occurs then the ratio increases to
greater than 1.
 Explain what
is flipped
ratio.
The ratio is used for

evaluation of patients with

50% MI: flipped ratio in 48hours
80% MI: flipped ratio in 72
the possible cardiovascular
hours
injury.
 WHAT IS THE CLINICAL SIGNIFICANCE OF
LD6?
LD6 is an isoenzyme of LD that probably
represents alcohol dehydrogenase. This band is
rarely detected in electrophoresis and migrates
cathodic to LDH-5. It can also metabolize lactate
and has been identified in the sera of severely ill
patients.
arteriosclerotic cardiovascular failure
hepatic congestion due to
cardiovascular disease
liver injury secondary to severe
circulatory
insufficiency
 HOW CAN LD ISOENZYMES BE UTILIZED IN
DIAGNOSING ACUTE MYOCARDIAL
INFRACTION?
LDH increases in the blood 6 to 12 hours after an acute MI,
peaks within 24 to 72 hours and normalizes within 8 to 14
days. In the past, a ratio of LDH1 to LDH2 greater than 1
was considered to be specific for an acute MI. The overall
sensitivity of LD for AMI approximates 96%, while the
specificity averages about 67%. Since it is not a specific
marker for cardiac myocytes, and its levels can also increase
in many other conditions, LDH is no longer used in the
diagnosis of myocardial infarction. Nowadays, the only usage
for LDH in the evaluation of acute MI is to differentiate acute
from subacute MI in patients with elevated troponin levels
and normal creatine kinase (CK) and CK-MB levels.
CASE
STUDY!
A 46 year-old woman with history of menometrorrhagia for 5–
6 years due to a voluminous uterine fibroid was admitted to
Department of Neuroscience, Reproductive Sciences and
Dentistry, School of Medicine, University of Naples Federico II,
Naples, Italy with fever (temp over 39 °C) and strong pelvic
pain. Transvaginal ultrasound (US) showed diffuse
fibromatosis and two evident uterine masses: the first was 53
× 57 mm, submucous, in fundus-anterior wall; the other was
97 × 70 mm, subserous, in isthmus-posterior wall. Despite the
size of the masses, no alarming features were observed.
CASE
STUDY!
In order to preserve pelvic stability, the woman
expressed the wish to avoid total hysterectomy;
therefore, a laparoscopic myomectomy was
considered. During hospitalization the patient
showed an inflammatory state (elevation of
fibrinogen and C-reactive protein) with
intermittent fever (not exceeding 38 °C). Blood
cultures were negative.
CASE
STUDY!
Several measurements of serum LDH total activity were
performed, showing normal or only slightly increased values,
with the highest peak of 304 U/l (reference range: 125–243).
Due to the clinical presentation, an abdominal CT with and
without contrast was performed, showing increased uterine
volume with two evident masses: the smaller one (4 × 3 cm)
was subserous, on the fundus; the larger one (12 × 10 cm),
voluminous and inhomogeneous, was para-uterine, on the
left, with intraligamentary growth and eccentric areas of
colliquative necrosis. These features were suggestive of
sarcomatous degeneration, so a MRI was recommended.
CASE
STUDY!
The abdomino-pelvic MRI was performed with and
without contrast, confirming the presence of both
masses (3.2 × 5 cm and 15x10x9 cm). In order to
support suspicion of malignancy, electrophoresis of LDH
isoenzymes was performed, showing the following
results: LDH1 = 15.6% (reference range: 16.1–31.5%);
LDH2 = 23.3% (29.2–41.6%); LDH3 = 21.0% (17.0–
26.2%); LDH4 = 13.6% (5.9–12.3%); LDH5 = 26.5%
(3.2–
17.3%); LDH5/LDH1 ratio = 1.7 (normal value: < 1).
CASE
STUDY!
Given the LDH isoenzymes pattern,
characteristic
supported by the CT report, a total abdominal
hysterectomy was performed in order to avoid
risks linked to the morcellation of
anoccult malignancy. Histological
examination of the surgical sample showed a
malignant mesenchymal proliferation constituted
by hypercellular areas with epithelioid or pleomorphic
cells alternating with hypocellular myxoid
areas. The definitive diagnosis wasof high
grade leiomyosarcoma
with myxoid changes, confirming suspicion.
DISCUSSIO
N1.What is the patient's diagnosis?
2.How did they come to this
diagnosis?
3.What is the relationship of LDH
isoenzymes to the patient's
diseased state?
QUESTIONS? THANK
CLARIFICAT ION YOU
S? FOR
LISTENIN
G!