MEDT 110- AUBF

SEMINAL FLUID- ADDITIONAL

TESTING

Sperm Vitality
Seminal Fluid
Fructose
Antibody- Antigen
in Seminal Fluid
Analysis

ADDITIONAL TESTING FOR ABNORMAL SEMEN ANALYSIS

ABNORMAL RESULT

POSSIBLE
ABNORMALITY

TEST

Decreased motility
with
normal count

Vitality

Eosin-nigrosin
stain

Decreased count

Lack of seminal vesicle

support medium

Fructose level

Decreased motility with

clumping

Male antisperm
antibodies

Mixed
agglutination
reaction and
immunobead tests
Sperm
agglutination with
male serum

Normal analysis with

continued infertility

Female antisperm
antibodies

Sperm
agglutination with
female serum

WHY is this test

requested?
TO KNOW THAT A MAN CAN
BECOME GENETIC FATHER
OR NOT

SPERM VITALITY

SPERM VITALITY
Sperm viability (vitality) should be
assessed if a low percentage of
sperm are progressively motile, e.g.,
30 - 40%.
Since motile cells are inherently
viable a viability assessment may not
be necessary when motility is high.
This test is important to determine if
the non-motile spermatozoa are alive
or dead.

SPERM VITALITY
Sperm vitality should be
assessed within 1 hour of
ejaculation.
Note: If <10% progressively motile
spermatozoa---do
vitality
assessment.

VITALITY TEST USING EOSINNIGROSIN

PRINCIPLE:
This one-step staining technique
uses nigrosin to increase contrast
between the background and the
sperm heads, which makes them
easier to discern. It also permits
slides to be stored for re-evaluation
and quality- control purposes.

VITALITY TEST USING EOSIN-NIGROSIN

METHOD:
1. 1 drop semen + 1 drop eosin
nigrosin
2.
Wait for 30 sec
3.
Put a drop on the slide
4.
Air dry
5.
Examine under oil immersion and
count at least 200 sperms
6.
Red sperms are not viable, white
sperms are viable

VITALITY TEST USING EOSINNIGROSIN

SEMINAL FLUID
FRUCTOSE

SEMINAL FLUID FRUCTOSE

Low sperm concentration may be
caused by lack of the support
medium produced in the seminal
vesicles, which can be indicated by a
low to absent fructose level in these
men.

SEMINAL FLUID FRUCTOSE

Low fructose levels are caused by:
abnormalities of the seminal
vesicles
bilateral congenital absence of the
vas deferens
obstruction of the ejaculatory duct
partial retrograde ejaculation
androgen deficiency.

SEMINAL FLUID FRUCTOSE

Specimens can be screened for the
presence of fructose using the
resorcinol test that produces an
orange color when fructose is
present.
A normal quantitative level of
fructose is equal to or greater than
13 mol per ejaculate.

SEMINAL FLUID FRUCTOSE

Specimens for fructose levels should
be tested within 2 hours of collection
or frozen to prevent fructolysis.

SEMINAL FLUID FRUCTOSE

Seminal Fructose Screening Test
1.Prepare reagent (50 mg resorcinol
in 33 mL concentrated HCl diluted
to 100 mL with water).
2.Mix 1 mL of semen with 9 mL of
reagent.
3.Boil
4.Observe for orange-red color

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
The results in the clumping and
inactivation of the sperm.
Antisperm antibodies can be present
in both men and women.

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
They may be detected in :

Semen
cervical mucosa
serum
and are considered a possible cause of
infertility.

It is not unusual for both partners to

demonstrate
antibodies,
although
male antisperm antibodies are more
frequently encountered.

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
Under normal conditions, the blood
testes barrier separates sperm from
the male immune system.
When this barrier is disrupted, as can
occur following surgery, vasectomy
reversal (vasovasostomy), trauma,
and infection, the antigens on the
sperm produce an immune response
that damages the sperm.

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
DEGREE OF
AGGLUTINATION
A. Head-to-head
B. Tail-to-tail (heads are
seen to be free and move
clear of agglutinates)
C. Tail-tip-to-head-tip
D. Mixed (clear head -tohead and tail -to- tail
agglutinations)
E. Tangle (heads and tails
enmeshed. Heads are not
clear of agglutinates as
they are in tail-to-tail
agglutination)

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
REPORTING on microscopic
examination

few
moderate
many

ANTIBODY- ANTIGEN IN SEMINAL

FLUID ANALYSIS
Two frequently used tests to detect
the presence of antibody-coated
sperm:

1. MIXED AGGLUTINATION
REACTION (MAR)
2.IMMUNOBEAD TEST

MIXED AGGLUTINATION REACTION

(MAR)
The MAR test is a screening
procedure used primarily to detect
the presence of immunoglobulin G
(IgG) antibodies.
The semen sample containing motile
sperm is incubated with IgG
antihumanglobulin (AHG) and a
suspension of latex particles or
treated RBCs coated with IgG.

MIXED AGGLUTINATION REACTION

(MAR)

MIXED AGGLUTINATION REACTION

(MAR)
In AHG combines with particles and
antibody-coated sperm forming
clumps.
Normal is <10% motile sperm attach to
particles

IMMUNOBEAD TEST
The immunobead test is a more
specific procedure in that it can be
used to detect the presence of IgG,
IgM, and IgA antibodies and
demonstrates what area of the
sperm
(head,
neckpiece,
midpiece,
or
tail)
the
auto
antibodies are affecting.

IMMUNOBEAD TEST
In the immunobead test:
sperm are mixed with
polyacrylamide beads known to be
coated with either anti-IgG, antiIgM, or anti-IgA.
Beads are coated with antihuman
globulin.

IMMUNOBEAD TEST
Depending on the type of beads
used, the test could be reported as
IgM tail antibodies,IgG head
antibodies, and so forth.

IMMUNOBEAD TEST
According to WHO:
Normal = 50% motile spermatozoa
with bound particles

OTHER TEST USED TO EVALUATE

MALE ANTISPERM ANTIBODIES
Gelatin agglutination tests
Double fluorochrome- sperm
cytotoxic antibody assay
ELISA
Mixed antiglobulin reaction

Opportunities are equal for all, but the

difference is --successful person gives
result ; unsuccessful person gives
only reasons.
(C)
Reported by:
JEVERLYN HERNANDEZMALIMBAN
BSMT 3-2
201313629