Non Invasive Prenatal
Testing
Presenter: Dr Amna Dilawar
Facilitator: Dr Raazia Tanveer
• NIPT is a test that uses maternal blood sample to determine if the baby has
certain chromosomal conditions, like Down syndrome and trisomies that can
effect health of the baby and pregnancy outcome.
• Includes Cell-free fetal DNA sequencing.
CELL FREE DNA
Both maternal and fetal DNA are found in maternal circulation.
ORIGIN:
Maternal cell free DNA : Hematopoietic cells apoptosis (long
fragments)
Fetal Cell free DNA : Placental cells (syncitiotrophoblast) apoptosis
Half life one hour. Cleared within 2 days of delivery.
• DNA is in small fragments.
• Highly fragmented each fragment between 50-200 base pairs.
• cfDNA represents the entire genome of mother and fetus
• Can be detected in maternal blood as early as 5 weeks.
• Almost always by 9 weeks.
• Comprises of 11-13% of total cfDNA in late first and early 2nd trimester.
• 0.1% rise per week till 20 weeks
• 1% per week rise from 20 weeks till term.
CLINICAL USE
ANEUPLOIDY DETECTION:
Secondary screening test is a non diagnostic follow up test offered to a
population that has already been found to be high risk. It is used to
detect DS, trisomies 18 and 13 in high risk population.
Fetal Rh-D typing:
Male fetal sex determination : For X-linked disorders like congenital
adrenal hyperplasia
Diagnosis of Single Gene disorder: eg. Achondroplasia
Methodology:
• Chromosome of Interest:
>> Most common method is counting of cfDNA fragments of
chromosome of interest. Mostly chromosome 21.
> It contains 1.3% of human genome
> Expected percentage of Chromosome 21 fragments in euploid
pregnancy is also 1.3%.
> In downs syndrome there is an extra 21 chromosome, the proportion
of fragments will be higher.
• Shotgun sequencing
• Targeted sequencing
• Possible test refinements
ACOG Screening
Recommendations:
Pre test counselling/Assess risk for aneuploidy
1.<35 year At least one of the following [Link]/Husband
1.>35 year at delivery Known carrier
3. Normal USG Of chr.r
2. Deranged serum markers abnormality
[Link]
significant [Link] abnormal USG finding 2. USG findings
family Hx not related to DS,
[Link] baby with Trisomy 18,13
aneuploidy
SCREENING
NIPT AMNIOCENTESIS/CVS
SERUM MARKERS
NIPT
NEGATIVE WITH NEGATIVE WITHOUT
POSITIVE
USG ANOMALIES USG-ANOMALIES
GENETIC COUNSELLING GENETIC Second trimester AFP
COUNSELLING R/o Neural tube defects
only
AMNIOCENTESIS/CVS AMNIOCENTESIS/
CVS/ DIAGNOSTIC
Microarray
Screening performance:
Most sensitive SCREENING TEST for following aneuploidies.
• Trisomy 21: DR 99.5%, FPR : 0.05%
• Trisomy 18: DR:97.7% FPR: 0.04%
• Trisomy 14: DR 96.1, FPR 0.06%
FALSE POSITIVE RESULTS:
• Confined placental mosaicism
• Demised Twin
• Maternal mosaicism
• Maternal Cancer
• Technical issue
• Recent blood transfusion
• Transplant recipient
FALSE NEGATIVE RESULTS
• Confined placental mosaicism
• Borderline low fetal fraction
• Maternal copy variants
• Technical issues
Causes of low fetal fraction:
• Early gestational age
• Suboptimal Sample collection
• Obesity
• Fetal Karyotype
PATIENT EDUCATION AND
COUNSELING:
• Done pre test and post test
1. Screening is optional
2. Limitations of screening vs diagnostic testing
3. Basic principles of cfDNA technology
4. DR, FPR, failure rates
5. Limitations of test, timings, incidental findings
6. Need to confirm abnormal results before termination
PROFESSIONAL PRACTICE
GUIDELINES:
• Sequencing of cfDNA is sensitive and specific for 21, 18, and 13
• Should be offered to hugh risk population
• Patient and provider education is important
• Insufficient data for twins
• Should not be offered to low risk until more info is available
• Obesity complicates the result :-P
Jazakillah
