Artifacts in Nuclear Medicine with Identifying and resolving artifacts.
screening prenatal test counseling in Hemoglobinopathies Thalasemia.pptx
1. Dr Ebin Roshan Paul
Screening, Prenatal diagnosis and
Counselling of Hemoglobinopathies
2. Screening for Hemoglobinopathies
Why screening important ?
1. Early detection and intervention of Thal.Major, severe
form of Thal.Intermedia, SCD will help to reduce the
mortality and morbidity of condition, also improves the
QoL.
2. Detection of carrier status will help to prevent birth of
an affected child in future.
3. Target population, Timing and
strategies
No single strategy can meet the need of population.
Population selection and timing of test is important will
determine options for future.
4.
5. Newborn screening
DBS spot card used / EDTA - heel prick/cord blood
4 good quality spots
Ideally within 24 hours of collection
With proper labelling place of birth and MR number
prior transfusion if any
family of origin and demographics
antenatal result if present
6. Techniques for Analysis
Primary Screening on DBS card
1. HPLC
2. Isoelectric Focusing (IEF)
3. Capillary Electrophoresis (CE)
A second line test needed on the same sample to validate
findings – protein sequence analysis ( mass spectrometry) or
DNA analysis
7. Screening is not a diagnostic test and not have 100%
sensitivity and specificity.
HPLC and IEF are the preferred methods for newborn
screening on the basis of cost-benefit analysis.
8.
9. Benefits of NB screening
Lethal complications can occur in presymptomatic age.
Early detection and comprehensive care mortality and
morbidity.
Prophylactic penicillin and PCV vaccination for SCD to
prevent pneumococcal sepsis.
10.
11. Childhood (6months-6years)
Universal screening for severe anaemia ( <7mg/dl )
Thal.Major and SCD can be identified, Thal.Intermedia not
picked up
Cost effective and since preschool children, approached
through Anganwadi.
12. Adolescence
Prior intense education programme before screening
Benefits: time to adapt with information, future partner
selection choices, helps to remove stigmatization.
Those who cannot avoid marriage with carrier prenatal
diagnostics and other choices.
Pilot project in Uttrakhand showed high acceptance and
retention of information by adolescence.
13. Premarital population
Not applicable to all society due to fear and stigmatisation.
Options provided are either avoiding the marriage or go
with prenatal diagnosis before each pregnancy.
Religious beliefs and local customs can also influence in
decision.
14. Prenatal Diagnosis
Carrier couples
First started by globin chain synthesis analysis in fetal blood
around 1970s by placental aspiration.
Biochemical and molecular analysis used for PND.
RDB and ARMS for mutation detection.
15. Fetal DNA sampling methods CVS (>11wks)
Amniocentesis (>16 wks)
Cordocentesis (18-20wks)
Pre-requisite are Pre-test counselling and written
informed consent of couple
Carrier status of both parents
Blood group of mother to prevent Rh
incompatibility if present.
16. Choice for “at risk couple”
Prenatal testing is a choice ( mutation studies of parents and
then analysing the fetus affected or not, then giving
counselling and option of termination of affected fetus)
When not feasible due to ethical and cultural issues
not to have children
adoption
in vitro fertilisation ( PGD ) and AI
17. Limitations of PND
No test are absolute ( 2% error )
Decision of termination of an already established
pregnancy is usually difficult and create emotional stress
Post test counselling is very important for emotional stress
in future.
18.
19. Antenatal population
Universal screening should be offered with CBC in first
antenatal visit in all levels.
MCV(<80), MCH(<27), relative high RBC count, normal RDW
HbA2 and HbF analysis advised
If carrier status noted, then husbands evaluated for “at risk
couples” and if detected PND based on Gestational age.
20. Cascade screening
Siblings and extended family of patients and carriers
Uttrakhand project found this more effective in screening
for carriers and detect “couple at risk”.
Some parents unwilling to communicate diagnosis and
misinformed relatives may stay away from affected family
so need proper counselling.
21.
22. Genetic counselling
An educational process that seeks to assist affected and/or
other at-risk individuals to understand the nature of the
genetic disorder, its transmission and the options available
in management and family planning.
(Kelly, 1986; Harper, 2004).
23. Counselling should be in simple/local language, removing
tensions, misconceptions, social stigma.
Taking a proper family history and pedigree.
Couples should be reassured of getting healthy baby after
prenatal intervention (at risk couples).
Information about procedure and its risk associated (pre-test
counselling)
24. Those who cannot choose termination due to religious and cultural
issues should be given options of PGD, artificial insemination with non
carrier, adoption, not to have children.
Nature of condition, predicted phenotype, severity, treatment options
should be discussed (post-test counselling).
Implication of carrier status and recurrence chances in subsequent
pregnancy (25:50:25) should be discussed.
25. Screening and risk assessment of family/relatives and HLA
matching if planning transplantation.
In “multi level consanguinity” risk assessment is difficult and
chances of having other recessive or dominant condition to co
exist is high.
Usual screening for hemoglobinopathies may miss detecting
these conditions. Should be included in counselling.
26. Beta Thalassemia
Thal.Intermedia wide range of spectrum ranging from
features of T.minor to T.major
To assess the predictable phenotype and severity
evaluation of α γ δ genes also necessary.
Ratio of α to non α chains (β/δ/γ) correlates with the
clinical phenotype severity. (α : non α chain)
27. α thal in β homozygous condition can have mild clinical
condition than β homozygous alone ( excess α chain ) and
decreases the severity.
Coexisting HPFH with β thal homozygous can have a milder
phenotype. Free α will go and bind with γ chain.
“δ/β double heterozygous” can have HbA2 normal and
resemble like α thal heterozygous.
“α/β double heterozygous” can have normal RBC indices
and elevated HbA2 level.
28. HbE disease (EE)
Counselling asymptomatic, no anaemia, no haemolysis,
no SM
Parental screening is mandatory for heterozygous status
(AE) and beta thalassemia status
Main differential diagnosis at birth is Hb Eβ0 but it will
symptomatic after the disappearance of HbF.
29. Hb E trait (AE)
1) Counsel baby will not have any health problems. Repeat
test advised at 6 month to confirm.
2) In future before marriage partner has to be screened for
hemoglobinopathies.
3) Parental screening is advised, to check beta thalassemia
status and assess future risk of having a HbE/Beta
thalassemia baby.
30. HbE-Beta Thal / HbE- β+ or β0
Wide range of phenotype from Thal major (severe) mild
forms of Thal Intermedia Thal minor (mild)
Majority will be moderate severity. Hb 6-7 g/dl. CF similar
to Thal Intermedia. Usually not need transfusion. Iron
overload may occur in future.
Coinheritance of α thal with HbE-β thalassemia result less
severity and milder phenotype. Reduces the α chain and
reduces the α : non α chain imbalance.
31. Coinheritance of determinants that increase HbF
expression can also decrease the severity of HbE-β
thalassemia.
Association of milder forms of β+ thalassemia (with
production of some β chain) are also have milder forms of
HbE-β thalassemia.
32. Hb SE
Compound heterozygous SE.
Mild chronic haemolytic anaemia, vaso-occlusive crisis are
rare. Usually presents at second decade of life.
Few cases reported all over world, 8 cases from south
India.
33. HbS-Beta Thal ( HbS / β+ or β0 )
compound heterozygous (βs/βthal)
Only few cases reported tribal population India.
Tribal areas: Asymptomatic or mild with a few vaso-occlusive
crisis. Most were having β0 mutation with associated α
thalassemia prevalence. (Mukherjee 2010)
Non tribal areas: noted severe β+ mutation and clinically more
severe than the tribal area. (Mukherjee 2010)
34. SCD – counselling dilemmas
India
Situation in India is different. SCD is prevalent mainly
scheduled caste and tribe population who are in low SES,
living in illiteracy, poverty and have local religious beliefs.
Need extensive counselling in local language, visual aids to
create awareness and need for testing.
Unlike β Thal the clinical severity is much less predictable in
SCD. India has a very variable range of clinical presentation.
Very few follow up data.
35. Presence of the Arab-Indian haplotype, higher HbF levels
and the presence of associated α-thalassemia mitigate the
severity of the disease in some tribal groups and it is not
always predictable. (Mukherjee 2000,2002)
Most often cases parents opt termination in homozygous
fetus there are chance that some may have a milder
phenotype and can live up to a reasonable age with
disease. (Colah 2005)
37. Various programmes and
Initiatives
Jai Vigyan programme : by ICMR covering 6 states
(Maharashtra, Gujarat, Karnataka, West Bengal, Punjab,
Assam). Beta thal carrier frequency 0 - 9.3%. Assam HbE
prevalence 41-66%.
West Bengal Thalassaemia Control Programme (WBTCP) :
2 nodal centres and 21 centres. Beta Thal prevalence 4 to
10%. Started prenatal analysis.
38. Sankalp India Foundation: Bangalore based NGO blood
donation, chelation aid, thalassemia relief, Bone marrow
transplant aid.
Gujarat TCP by Red Cross Society: screening in college and
schools. Started PND centres.
Maharashtra: 4 district started screening, prevention and
treatment for Thalassemia and SCD.
39. Punjab : “Project Rainbow” ( Center, State, NGO,
Corporate, patients-parents groups ) for care of
Thalassemia patients.
“ThalCare” : Web based platform by Sankalp Foundation
for thalassemia management. Used here
“Thalaman”: software developed to store and assimilate
data online.
40. “Thalassochip” : diagnostic tool for beta thalassemia based
on aaryed primer extension ( APEX). Help to identify beta
thal mutation and Hb variants on a low operating cost.
41. Sickle cell anaemia control programme (SCACP)
Gujarat : started in 2006, mainly targeting tribal
population. All antenatal are given screening on
“Mamta Divas” – a special immunisation day, if found
carriers then father is screened and given counselling.
Colour coded cards are given for people based on
status.
42. Road map ahead.....
Need of education and awareness generation in rural and
urban areas of various states.
Most of data from hospital and selected population not
reflect actual disease burden. Several states and ethnic
groups, community not screened and “micro mapping”
needed in every state.
43. Adequate centres, trained staffs and counsellors need.
Providing hands on training at regular intervals.
Establishment of more day care centres as patient load
increasing.
Encouragement of bone marrow transplant in low risk
patients and affordable families. Establishment in
government set up.
44. Joint and sincere effort from Center, State, Various NGO’s ,
Corporate houses, patient support groups backed by
political support for the implementation of the National
Control Programme.