Paternity Testing - Six different red blood cell antigen are commonly used in parentage :

- Paternity test now available can prove with nearly 100% certainty the - The ABO blood group system : is the first system of antigen to be tested
father of the almost any child, even before birth. in paternity testing
- The assessment of paternity may be necessary in a variety of cases - Although the use of A and B antigen markers alone is of limited value, it
including divorce, custody case and charges of rape and incest. may demonstrate a definitive exclusion,
- Establishment of parentage may also be desirable to authenticate a - If an exclusion cannot be confirmed, antigens of the remaining five
child’s medical history or personal identity principal red blood cell systems should be tested.
- If parentage is uncertain, blood test can be used to identify specific - Because red blood cell antigen testing usually yields a relatively low
genetic markers. probability of excluding a falsely accused male, it must be often used in
- Genetic markers can be : conjunction with other genetic testing system
o Red blood cell antigen
RBC Antigen Systems Antigens
o Red blood cell enzymes
ABO A, A1, B
o Serum proteins
Rh D, C, c, E, e
o Human Leukocyte Antigen (HLA) MNS M, N, S, s
o DNA analysis Kell K, k, Kpa, Kpb
- The statistical probability of alleged paternity can be established on the Duffy Fya, Fyb
basis of the combined frequency of genetic markers Kidd Jka, Jkb
- Exclusion of paternity can be achieved with relative certainty
- Genetic markers used in parentage testing must have certain Red Blood Cell Enzymes
characteristics in order to be useful in parentage assessment. - Some red blood cell enzymes can be used in parentage assessment
- Fundamental characteristics includes: - These enzymes exhibit genetic polymorphism
o A pattern of Mendelian Inheretance - The usefulness of RBC enzymes is limited because of technical difficulties
o Low frequency mutation and recombination o Accuracy
o Reproducibility
Characteristics if Genetic Markers in Parentage Testing - Enzymes used are :
1. Exhibit a pattern of Mendelian dominance o Acid Phosphatase
2. Have low frequency of mutation and recombination o Adenosine deaminase
3. Are adequately developed at birth or early delivery o Adenylate Kinase 1
4. Persist throughout a person’s lifetime without environmental or o Esterase O
genetic alteration o Glucose 6 Phosphate Dehydrogenase
5. Are demonstrable and reproducible in the laboratory o Glycoaminase 1
o Phosphogluco__________ 1
o Phosphogluco __________ 2
o 6-phosphoglucose dehydrogenase

Red Blood Cell Antigen

Serum Protein Polymorphism Deoxynucleic Acid (DNA) Analysis
- Some serum proteins have been shown to be disseminated on as - DNA analysis has become increasingly common because of its standing
inherited basis. accuracy.
- These proteins are multi-molecular in form and can be demonstrated in - This relatively new technology has applicability in paternity testing
the laboratory - More than 30 DNA probes are currently available for investigating
- Because serum protein polymorphism measurements are very labor virtually any biologic parentage relationship
intensive, they are usually used in adjunct testing. - To prepare DNA for study using a DNA probe, the DNA is cut into
- In addition, a child must be at least 6 months old for most testing ( for fragments using the restriction fragment length polymorphism (RFLP)
example : Gm allotyping) test.
- This eliminates the effect of the large amounts of maternal IgG - DNA is removed from cells usually taken from blood samples and
circulating on the child which may cause direct exclusion to be missed. precisely cut apart at specific sequences using specialized restrictions co-
- Indirect exclusions are valid only when the phenotype differs that of the enzyme that act as biologic scissors.
mother. - After electrophoresis (separation of proteins through the use of an
- Km allotypes are not age-dependent electrical current), the bands of DNA fragments are transferred from the
- Serum proteins used are : gel medium of the test onto a nylon membrane.
o Gm protein - The membrane is heated and gene probe labelled with a small amount of
o Complement (C3) radioactive material added.
o Haptoglobin - A gene probe is a small DNA segment manufactured in the laboratory
o Transferrin and tagged with radioactive marker.
- If the probe and the subject’s DNA piece match, the probe displays the
Human Leukocyte Antigen subject polymorphic DNA position on the membrane by emitting
- The Major Histocompatibility Complex MHC in man is termed as Human radioactivity onto the film.
Leukocyte Antigen System - This activity produces an x-ray like picture, an autoradiogram of the DNA
- HLA antigens are expressed on almost every type of nucleated cell of the mother, the child, and the alleged father.
- Lymphocytes have the highest density of expressed antigens - In the future, another method based on polymerase chain reaction (PCR)
- HLA used are : amplification may replace conventional analysis.
o HLA-A - This technique, amplified fragment length polymorphism (AMPFLP)
o HLA-B produces millions of copies of only the specific portion of the
o HLA-C chromosomes containing the fragment length polymorphism.
o HLA-D o This method is faster than traditional analysis, requires smaller
o HLA-DR blood samples, and works on aged blood samples.
o It is also less labor intensive.
Determination of Paternity  Cumulative probability of paternity results from the
- In order to be able to determine exclusion or non-exclusion, the results multiplication of W for each genetic system is tested.
of the laboratory testing must be analysed as to the identity of the
obligatory paternal genes.
- Obligatory paternal genes are those genes that the child must inherit
from the biological father.
- The genetic systems currently in use have a cumulative power of
exclusion (P) of greater than 95%.
- Little difficulty should be encountered in excluding an alleged father.
- Two types of exclusion exist:
o 1st order
o 2nd order
- A first order exclusion is one in which the child has a gene that both the
mother and alleged father lack.
- Second order exclusion occur when the genes present in an alleged
father are not represented in the child – that is, the paternal obligatory
genes are not found in the alleged father bit are present in the biological
mother.
- When an exclusion cannot be found, the results of the genetic testing are
used to calculate the combined paternity index (PI) and/or cumulative
probability of paternity (W).
- The PI is a ratio that compares the relative probability of paternity of the
putative father to that of a random male for any given genetic system
- Illustration : PI = X/Y
o X : the frequency with which the alleged father (af) passes on
that obligatory gene
o Y : the frequence with which the random male (rm) passes on
that obligatory gene.
o The combined paternity index is achieved by multiplying the PI
for each genetic system tested
- The probability of Paternity (W)
o Is the percent chance that the alleged father is the biological
father based on the frequency of the genetic markers detected.
o The statistical formulas used for this calculations are predicted
using Bayer Theorem : where a prior probability of 0.5 is
assumed.
o Illustration : W – x(x/y)