Seminal stains.

Q. 1 Best test for seminal stains is:

 A Barbeiros test

 B Acid Phosphatase test

 C Florence test

 D Alkaline Phosphatase test

Ans. B

Explanation:

The acid phosphatase test- 1. Level > 25 kA Unit is specific for the test or Undiluted
semen has an acid phosphatase activity of 340 – 360 Bodansky U/ml, it is higher in

humans as compared to animals. 2. It is +ve also by human RBCs, semen of higher

apes and juice of cauliflower. 3. The prostatic secretion element of seminal fluid
contains a very much higher percentage of acid phosphatase. 4. This test is

conclusive in the absence of demonstrable sperms or in aspermia. 5. Is a

quantative test.

Q. 2
Dried semen stain in clothes is identi ed by

 A UV light

 B Spectroscopy

 C Magnifying lens

 D Infra red
Ans. A

Explanation:

UV light [ Ref: Reddy 29/e, p 377-378; Parikh 6/e, p 7.26-7.271

Seminal stains have to be detected in sexual offences. These have to be
differentiated from those due to starch, pus, vaginal discharge, urine & egg

albumen.
Seminal stains may be examined by four methods – (1) Physical (2) chemical
(3) Microscopic (4) Electrophoretic

1) Physical examination – A preliminary examination under filtered ultraviolet light

can be made. Stains of urine, vaginal discharge and seminal fluid show a bright
fluorescence. The fluorescence of seminal stains is of a bluish white colour & such

stains are selected for further examinations.

2) Chemical examination – Following tests can be done

Florence test – It is based o the presence of choline in semen. After adding

reagent, dark brown crystals are formed.

Barberio’s test – It depends upon the presence of spermine in semen. Yellow,

needle shaped, rhombic crystals of spermine pictrate are produced when

reagent is added.

The acid phosphatase test – A high concentration of acid phosphatase is found

only in the semen of human beings & monkeys. This is a quantitative test. The

amount of acid phosphatase is estimated. Inhibition of acid phosphatase by 1-

tartaric acid, though non specific, is a valuable screening test.

Creatine phosphokinase test – Spermatozoa contain a high concentration of

cratine phosphokinase. Levels over 400 U/ml is almost diagnostic of semen.

The enzyme is stable and can be detected even in old stains of 6 months.

Choline and spermine test – Liquid semen and dried seminal stains can be

identified by a thin layer chromatographic technique. The test is based on the

unique combination of choline & spermine which is present only in semen.

Ammonium molybdate test – when reagent is added to stain, a deep yellow

colour from the presence of phosphorus is produced.

Immunological method – MHS-5 from seminal vesicles and Mab 4 E6 can be

detected on the sperm cells & in ejaculated fluid. Prostate specific antigen
 (PSA, P30) test has replaced the quantitative analysis for acid phosphatase

[Fluroscent in situ hybridisation (FISH) has been suggested as a specific &

sensitive test for detection of male epithelial cells in the post coital vagina upto

one week]

3) Microscopic examination – Involves identification of spermatozoa under the

microscope. Fluorescence microscopy is based on the principle that the Y-

chromosome is fluorescent to quinacrine.

4) Electrophoretic methods
Acid phosphatase isoenzyme test – Polyacrylamide gel electrophoresis followed

by staining with methyl umbelliferyl phosphate reagent makes the seminal acid

phosphatase to be distinguished from the acid phosphatase present in other

substances & in vaginal secretion.

This is superior to LDH isoenzyme method since semen can be identified in the

absence of sperms as in azoospermic or vasectomised persons.

Lactate dehydrogenase (LDH) isoenzyme Test – Sperm specific LDH isoenzyme

can be separated from other LDH isoenzymes of semen by polyacrylamide gel

electrophoresis.

Proof of semen: The only absolute proof of semen is the finding of at least one
unbroken spermatozoan, or electrophoretic LHD isoenzyme detection of sperms.

In case of azoospermia, vasectomy or very old persons a stain will be seminal

if it gives

– characteristic fluorescence in UV light

– positive 1-tartarate acid-phosphatase and

– High levels of acid phosphatase & creatine phosphokinase

or electrophoretic seminal acid phosphatase or PSA (P30) positive

Species (Human or animal) is confirmed by precipitin test as in blood stains.

Source (from which person) is identified by DNA & different grouping systems

such as ABO, GM, PGM & GLO-I. New test

Commercial ELISA, the SEMA®assay, for a seminal vesicle-specific antigen

(SVSA) provides highly sensitive detection of semen but are costly .

Q. 3 Which is a not an essential criteria according to WHO for normal semen analysis

 A Sperm count > 20 million/ml

 B Volume > 1 ml

 C Sperm with normal morphology (strict criteria) > 15%

 D Motility > 25% with rapidly progressive motility

Ans. B

Explanation:

Volume > 1 ml [Ref William’s Gynaecology p. 449;

http//www.gfiner.ch/end%•pgi/network]

THE W.H.O. RECOMMENDS THE FOLLOWING NORMAL VALUES

Volume > 2 ml
Sperm concentration > 20 million / ml or more
Total sperm number > 40 million/ejaculate
Percent mobility > 50% with forward progression
Forward progression > 25% with rapid linear progression
Normal morphology > 2 (scale 0-4)
Round cells > 30% normal (W.H.O. criteria)
Sperm agglutination > 14% normal (Kruger and strict criteria)
< 5 million/rnl
< 2 (scale 0-3),w.b.c fewer than I x106 / ml

The normal sperm morphology is currently being reassessed by W.H.O

In the interim the proportion of normal forms accepted by the laboratories in

the U.K is either the earlier W.H.O limits of 30 or 15% based on strict

morphological criteria.

Q. 4
Color of dried semen becomes yellow to brown. Dried semen stain in clothes is
identi ed by:

 A UV light

 B Spectroscopy

 C Magnifying lens

 D Infrared

Ans. A

Explanation:

When dried semen is examined under filtered ultraviolet light, they show a

fluorescence of a bluish-white colour, which is not specific as other albuminous

materials such as nasal, leucorrhoeal discharges and detergents also fluoresce.

Fluorescence depends on choline of semen.

Ref: The Essentials of Forensic Medicine and Toxicology, 27th edition, Page 369.

Q. 5
Which of following tests in used to detect semen?

 A Phenolphthalein test

 B Reine’s test

 C Barberio’s test

 D Para n test

Ans. C

Explanation:

C i.e. Barberio’s test :

 Barberio’s test is used for identification of seminal stains

Few drops of barberio’s reagent (containing pieric acid) are added to the seminal
stain.

Positive test is indicated by formation of Yellow and needle shaped crystals of

spermine picrate.

(Spermine in seminal stain reacts with picric acid to form these crystals)
Phenolphthalein test (Kastle-Meyer test) is used for identification of

haemoglobin in blood stains – Parikh 6th/7.16

Paraffin test (or Dermal nitrate test) is used to detect gun powder on skin i.e.

to determine if suspect had discharged a firearm- Parikh 6″/7.39

Reine’s test has not been mentioned in forensic literature

Forensic tests to detect

Seminal Stain Blood Stains

– Barberio’s testO – Benzidine testQ

– Fluorence testQ – Phenophthalein (Kastle‑

– Acid phosphatase Meyer) testQ

testQ – Takayama’s

– Creatine phosphatase Haemochromogen crystal

test testQ

– Elisa test – Teichmann’s Haemin crystal

testQ

– Spectroscopic test

Q. 6
Spermin is detected in semen by which test‑

 A Barberio test

 B Florence

 C ELISA
  D Agglutination inhibition

Ans. A

Explanation:

A i.e. Barberio test

Q. 7
All are tests for seminal stains except:           
March 2012

 A Florence test

 B Creatine phosphate test

 C Takayama test

 D Barberio test

Ans. C

Explanation:

Ans: C i.e. Takayama test

Stains and features

If semen is present, florence test will show dark brown crystals of choline
iodide.

Levels over 400 units of CPK/ ml are almost diagnostic of seminal stains

Blood stains with Takayama reagent produces pink, feathery crystals of

haemochromogen/ reduced alkaline haematin.

If prostatic secretion is present, yellow needle shaped rhombic crystals

(spermine picrate) are seen

Q. 8
Florence test is used for: 
Delhi 06

 A Blood stains

 B Seminal stains

 C Salivary stains

 D Sweat stains

Ans. B

Explanation:

Ans. Seminal stains

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