F1: Semen Analysis (Part 4: Tests for Semen Analysis) Flashcards
- Done in cases where the sperm count is low
- Evaluates capacity of seminal vesicle to provide fructose to sperm
- Based on the principle of Resorcinol test
Seminal Fluid Fructose
Used as nutrition for sperm
Fructose
Specimens for fructose levels should be tested within how many hours to prevent what? If it cannot be tested within that time, what should be done?
2 hours of collection to prevent fructolysis (or frozen)
Positive result for Resorcinol Test?
Red Orange
Normal quantitative level of fructose?
Equal to or greater than 13 µmol per ejaculate
High or Low Fructose Levels?
- Abnormalities of the seminal vesicles
- Bilateral congenital absence of the vas deferens
- Obstruction of the ejaculatory duct
- Androgen deficiency
- Partial retrograde ejaculation
Low Fructose Levels
Phenomenon when the semen travels backwards into the bladder
Retrograde Ejaculation
- Uses Florence reagent (Iodine crystals and KI)
Florence Test
Composition of Florence reagent?
Iodine crystals and KI
Positive results of Florence Test?
Brown rhombic crystal (periodide of choline)
- Uses TCA and picric acid
Barbiero’s Test
Positive Results for Barbiero’s Test?
Yellow leaf-like crystals (spermine picrate)
- Test that utilizes a leukocyte esterase reagent strip that is used as screen for the presence of leukocytes
Leukocytes
- May be present in both men and women
- May be detected in semen, cervical mucosa, or serum
- Considered a possible cause of infertility
Anti-Sperm Antibodies
T or F: It is unusual for both partners to demonstrate antibodies, although male antisperm antibodies are more frequently encountered.
F (not unusual)
- Must be intact to prevent sensitizing the immune system from producing antibodies against own sperm
- Under normal conditions, this separates sperm from the male immune system
Blood Testes Barrier
When the blood testes barrier is disrupted, such as in surgery, vasectomy reversal or vasovasostomy, trauma, and infection, what happens?
The antigens on the sperm produce an immune response that damages the sperm.
T or F: The damaged sperm may cause the production of antibodies in the female partner.
T
Female or Male Sperm Antibody?
- You note clumps of sperm adhering to one another and poor motility
Male Sperm Antibodies
Female or Male Sperm Antibody?
- No clumps are seen
- After coitus, you examine cervical mucosa and you note clumps of sperm and poor motility
Female Sperm Antibodies
These are two frequently used tests to detect the presence of antibody-coated sperm.
- Mixed Agglutination Reaction (MAR)
- Immunobead Test
- Screening procedure to detect the presence of immunoglobulin G (IgG) antibodies
- The semen sample containing motile sperm is incubated with IgG antihuman globulin (AHG) and a suspension of latex particles or treated RBCs coated with IgG.
- The bivalent AHG binds simultaneously to both the antibody on the sperm and the antibody on the latex particles or RBCs, forming microscopically visible clumps of sperm and particles or cells.
Mixed Agglutination Reaction (MAR)
T or F: In MAR, a finding of more than 10% of the motile sperm attached to the particles is considered normal.
F (less than 10%)
Positive result for Mixed Agglutination Reaction?
Clumps of sperm and particles or cells
- A more specific procedure that can be used to detect the
presence of IgG, IgM, and IgA antibodies - it demonstrates what area of the sperm (head, neckpiece,
midpiece, or tail) the autoantibodies are affecting - Sperm are mixed with polyacrylamide beads known to be coated with either anti-IgG, anti-IgM, or anti IgA
Immunobead Test
Can interfere with penetration into the
cervical mucosa or ovum.
Head-directed Antibodies
Affect movement through the cervical
mucosa
Tail-directed Antibodies
Microscopic examination of the sperm in Immunobead Test shows what?
The beads attached to sperm at particular areas
What are the ways that Immunobead Test results may be reported?
These depend on the type of beads used.
IgM tail antibodies, IgG head antibodies, etc.
T or F: The presence of beads on less than 50% of the sperm is
considered normal as defined by the WHO.
T
Identify the test.
- Abnormal Result: Decreased motility with normal count
- Possible Abnormality: Vitality
Eosin-Nigrosin Stain
Identify the test.
- Abnormal Result: Decreased count
- Possible Abnormality: Lack of seminal vesicle support medium
Fructose Level
Identify the test.
- Abnormal Result: Decreased motility with clumping
- Possible Abnormality: Male anti-sperm antibodies
- Mixed Agglutination Reaction and Immunobead Test
- Sperm Agglutination with Male Serum
Identify the test.
- Abnormal Result: Normal analysis with continued infertility
- Possible Abnormality: Female anti-sperm antibodies
Sperm Agglutination with Female Serum
T or F: The presence of more than 1000 WBC per millimeter is
significant which indicates infection within the reproductive system, frequently the prostate.
F (more than 1 million)
What part of the reproductive system is most frequently infected?
Prostate
What organisms usually cause the infection of the reproductive system (frequently the prostate)?
- Chlamydia trachomatis
- Mycoplasma hominis
- Ureaplasma urealyticum
Sperm Function Test
- Observed under microscope, evaluates sperms penetration
Hamster Egg Penetration
Sperm Function Test
- Evaluates sperm penetration in partner’s cervical mucus (mid-cycle)
Cervical Mucus Penetration
Sperm Function Test
- Low sodium concentration
- Evaluates sperm’s membrane integrity & sperm viability
Hypo-osmotic Swelling Test
Sperm Function Test
- Evaluates acrosome to produce enzymes for ovum penetration
In-Vitro Acrosome Reaction
What are the 4 ways of detection of semen in medico legal cases?
- Microscopic Exam for presence of sperm cells (specimen with xylene and examining under phase microscopy)
- Prostatic acid phosphatase
- Seminal glycoprotein p30 (prostatic specific antigen [PSA]
- ABO blood grouping & DNA analysis
- A much less involved procedure when compared with infertility analysis because the only concern is the presence or absence of spermatozoa
- Recommended testing includes examining a wet preparation using phase microscopy for the presence of motile and nonmotile sperm
Post-Vasectomy Evaluation
T or F: The length of time required for complete sterilization can vary greatly among patients and depends on only the time of ejaculation.
F (it depends on both the number and time)
T or F: Finding viable sperm in a post-vasectomy patient is not uncommon, and care should be taken not to overlook a single sperm.
T
What shows a successful vasectomy?
- Specimens are routinely tested at monthly intervals, beginning at 2 months post vasectomy and continuing until two consecutive monthly specimens show no spermatozoa
If patient is negative for a post-vasectomy evaluation, what has to be done?
Centrifuge for 10 minutes then re-examine
- Measures the quality of cervical mucus and the ability of the sperm cells to penetrate the mucus and maintain activity
Sim Huhner Test (Post Coital Test)
Normal value for Sim Huhner Test?
Presence of 10 motile sperm cells/hpf 6-8 hours after coitus
- Test for the tenacity of cervical mucus
Spinnbarkeit Test
What is considered good tenacity for the Spinnbarkeit Test?
10 cm before breaking
Chemical Testing
(Increased/Decreased) level of fructose means lack support medium produced in the seminal vesicle.
Decreased
Chemical Testing
Decreased level of these indicate a disorder of the epididymis
Three answers.
- Neutral A glucosidase
- Glycerophosphocholine
- L-carnitine
Chemical Testing
Decreased level of these indicate lack of prostatic fluid
Two answers.
- Zinc
- Citric Acid Glutamyl Transpeptidase Acid Phosphatase
