Noninvasive prenatal testing (NIPT) has become possible to identify cell free fetal DNA (cffDNA) in maternal bloodstream. The effective implementation of noninvasive testing into a clinical practice such as fetal sex detection, RhD genotyping and fetal chromosomal aneuploidy detection have been enabled by the technical advances in molecular analysis of fetal DNA (e.g., digital polymerase chain reaction (PCR) and massively parallel sequencing (MPS) in early years. cffDNA technology has significant superiority as difference compared to routine serum sample screening by means of it has high accuracy and sensitivity. Though, cffDNA positive results still need validation by the invasive testing. The main purpose of cell free fetal DNA is to become first priority of NIPT option only due to its high accuracy rate and its safety. The ultimate goal is to develop the reliable method which could eventually swap invasive prenatal testing procedures. In maternal circulation with relatively high concentration the analysis of cell-free fetal DNA in plasma is one of the innovative non-invasive prenatal testing options. Recently Commercial tests for cell-free fetal DNA in maternal blood have become available. This review briefly summarizes the technical aspects of the noninvasive prenatal testing along with analysis of cffDNA and application of it in clinical practice.

1. Cell Free Fetal DNA
Non-invasive Prenatal Diagnostic methods
and Applications
SNJB’S Shriman Suresh Dada Jain College of Pharmacy, Chandwad
Presenter – Mayuri N Jagtap
Department- Pharmacology
SEMINAR

2. Prenatal
CffDNA
Noninvasive

3. CONTENT
ADVANTAGES OF NIPT
INTRODUCTION
DISADVANTAGES OF NIPT
DEVELOPMENT IN CFFDNA ISOLATION METHODS
ANALYSIS OF Cell FREE FETAL DNA
APPLICATIONS OF CELL FREE FETAL DNA
CONCLUSION
REFERENCE

4. INTRODUCTION

5. NIPT is based on Cff DNA known
as prime progress and does not
require supplementary testing
To reduce risk of invasive techniques
efforts has been made to emerge more
precise, dependable harmless NIPT i.e.,
cffDNA
It has high detection rate (~95%) and
has low false positive rate (~1%)
Cell free fetal DNA has made in
1997
The use of noninvasive testing in maternal
circulation at a high risk have been approved by
American congress of obstetrician & gynecologist
in 2012
Maternal plasma is mixture of maternal and fetal
DNA. Fetal DNA is shorter than the maternal
DNA

6. ADVANTAGES OF
NIPT

7. -cffDNA is definite to pregnancy
Reduced risk to the fetus
Involves only maternal blood test
It has high detection rate
Useful in gender determination
Available from 10 weeks gestation onwards useful for
those who have missed CFTS (combined first trimester
screening

8. DISADVANTAGES
OF NIPT

9. Anatomical abnormalities not detected by cffDNA
It is important to repeat that NIPT is a sensitive screening test
due to false positive and false negative results
Some type of NIPT not suitable for twins
NIPT is not a diagnostic test
It is impotent to discriminate between fetus and placenta by
means fetal DNA fragments are originate from placenta
It is not available everywhere & has false positive result for
cancer

10. DEVELOPMENT IN
CFFDNA ISOLATION
METHODS

11. The yield of cell free fetal DNA depends heavily on the isolation method
To follow fetal genotyping analysis in NIPAT it is necessary to choose the
optimal isolation process
The biggest challenge in cffDNA based NIPAT is interference from maternal
DNA, compare to conventional post natal paternity testing

12. Development in cffDNA
isolation methods
Extraction Method
Column
based DNA
Extraction
Magnetic
Extraction
Enrichment Method
Size
selective
enrichment
DMR based
enrichment
Formaldehyde
Based
enrichment

13. ANALYSIS OF
CFFDNA

14. Analysis of cffDNA do not isolate fetal & maternally detected
DNA but useful cffDNA supplement & by using automated data
processing tools, it determines the complete variation between
sequenced DNA fragments
The cffDNA Science testing have a
tendency to be very specific & extremely
reactive
The promising findings of cffDNA also needs
verified testing because of its protection & excellent
precision, the cffDNA analysis is prime alternative
for NIPT
The main aim is to establish a
rational system to interchange
invasive prenatal therapy

15. Analysis of
cffDNA
Shotgun
sequencing
Mass
spectrometry
Polymerase
chain reaction

16. APPLICATONS OF
CFFDNA

17. Prenatal Sex Discernment
Congenital adrenal hyperplasia
Paternity testing
Detection of maternal health
problems
Single gene disorder
Haemolytic disease of the
fetus & new-born
Aneuploidy
Preeclampsia

18. CONCLUSION

19. The main purpose of cffDNA is to
become first priority of NIPT
option only due to its high accuracy
rate.
Assignment summarizes the
technical aspects of the noninvasive
prenatal testing along with analysis
of cffDNA & application of it in
clinical practice.
The ultimate goal is to develop the
reliable method which could
eventually swap invasive prenatal
testing procedures.
In maternal circulation the analysis of
cffDNA in plasma is one of the
innovative non invasive prenatal
testing option.
Recently, commercial tests for
cffDNA in maternal blood have
become available.

20. REFERENCES

21. 1. Lo YM, Corbetta N, Chamberlain PF, et al, Presence of Fetal DNA in
Maternal Plasma and serum, 1997; 350(9076): 485-487
2. Wataganara T, Bianchi DW, Fetal Cell-Free Nucleic Acids in the
Maternal Circulation: New Clinical Applications, Annals of the New
York Academy of Sciences, 2004; 1022: 90-99
3. Sharma VK, Paul V, Glick J, Mass Spectrometric Based Analysis,
Characterization and Applications of Circulating Cell Free DNA
Isolated From Human Body Fluids, The International Journal of Mass
Spectrometry, 2011; 304(2-3): 172–183
4. Mayuri Jagtap, Manasi Gangurde, Cell Free Fetal DNA: Noninvasive
Prenatal Diagnostic Methods and Applications, International Journal of
Pharmaceutical Research, Jan - Jun 2020 | Supplementary Issue 1

22. THANK YOU
🐾