Preimplantation Genetic Testing (PGT)

Is a procedure used in conjunction with in vitro fertilization (IVF) to screen embryos for genetic
abnormalities before they are implanted into the uterus. This testing helps ensure that only embryos
without specific genetic issues are selected for implantation, thereby reducing the risk of certain genetic
disorders.

Types of Preimplantation Genetic Testing

[Link]-A (Aneuploidy):
Screens for chromosomal abnormalities, such as an incorrect number of chromosomes, which
can lead to conditions like Down syndrome.

[Link]-M (Monogenic/Single Gene Disorders):

Targets specific genetic disorders caused by mutations in a single gene, such as cystic fibrosis,
sickle cell anemia, or Huntington’s disease.

[Link]-SR (Structural Rearrangements):

Used to detect structural rearrangements of chromosomes, like translocations or inversions,
which can lead to miscarriages or congenital disabilities.

Procedure

•IVF Process: Embryos are created through IVF, where eggs are fertilized by sperm in a lab.

•Biopsy: A few cells are taken from the embryo (typically at the blastocyst stage) without harming its
development.

•Genetic Analysis: The sampled cells are analyzed for genetic abnormalities.

•Embryo Selection: Based on the results, only embryos without the detected abnormalities are selected
for implantation.

Benefits

•Reduced Risk: Lowers the risk of passing on genetic disorders.

•Higher Success Rates: Increases the chances of a successful pregnancy and healthy baby.
•Informed Decision-Making: Provides valuable information for couples at risk for genetic disorders

PGT is especially recommended for couples with a known risk of genetic disorders, repeated IVF failures,
recurrent miscarriages, or advanced maternal age.
Techniques Used In PGT:

1. Fluorescent In Situ Hybridization (FISH)

- **Purpose:** Detects chromosomal abnormalities, particularly aneuploidies (abnormal number of
chromosomes).
- **Method:** Fluorescent probes that bind to specific chromosome regions are used to visualize the
number and structure of chromosomes under a microscope.
- **Application:** Commonly used in PGT-A for detecting conditions like Down syndrome (trisomy 21)
but is limited by the number of chromosomes it can assess simultaneously.

2. Polymerase Chain Reaction (PCR)

- **Purpose:** Amplifies small DNA segments to detect specific mutations in single-gene disorders.
- **Method:** DNA from the embryo is extracted and amplified through PCR, allowing for the
identification of specific gene mutations.
- **Application:** Used in PGT-M for conditions like cystic fibrosis, sickle cell anemia, or Huntington’s
disease.

3. Comparative Genomic Hybridization (CGH)

- **Purpose:** Detects unbalanced chromosomal abnormalities, including duplications or deletions of
chromosomal segments.
- **Method:** DNA from the embryo and a normal reference sample are labeled with different
fluorescent dyes and hybridized to a microarray. Differences in the fluorescence ratio reveal
chromosomal imbalances.
- **Application:** Used in PGT-A and PGT-SR to assess the overall chromosomal balance in embryos.

4. Next-Generation Sequencing (NGS)

- **Purpose:** Provides a comprehensive analysis of genetic material, allowing for the detection of
chromosomal abnormalities, single-gene mutations, and even low-level mosaicism.
- **Method:** DNA is extracted from the embryo, fragmented, and sequenced. NGS can generate
massive amounts of data, which are then analyzed to detect genetic anomalies.
- **Application:** Used in PGT-A, PGT-M, and PGT-SR for detailed and accurate genetic screening.

5. Single Nucleotide Polymorphism (SNP) Array

- **Purpose:** Detects chromosomal abnormalities and can also be used for parentage testing,
identifying inherited genetic traits.
- **Method:** This microarray-based technique assesses variations at specific points (SNPs) in the
genome, comparing the embryo’s genetic profile with that of the parents.
- **Application:** Used in PGT-A and PGT-M to detect chromosomal imbalances and single-gene
disorders.

6. Array Comparative Genomic Hybridization (aCGH)

- **Purpose:** Similar to CGH but more advanced, used to detect more subtle chromosomal
imbalances.
 - **Method:** Uses a microarray platform to compare the genetic material of the embryo to a
reference sample, identifying gains or losses in chromosome segments.
- **Application:** Used primarily in PGT-A to detect aneuploidies and other chromosomal
abnormalities.

7. Karyomapping
- **Purpose:** Identifies chromosomal abnormalities and inherited genetic diseases by analyzing the
genetic markers across the entire genome.
- **Method:** Involves comparing the genetic markers (SNPs) of the embryo with those of the parents
and a close relative (usually a sibling) to detect the inheritance of specific chromosomal regions.
- **Application:** Used in PGT-M for single-gene disorders and chromosomal abnormalities,
particularly when the family history is known.

8. Quantitative PCR (qPCR)

- **Purpose:** Allows for the quantification of DNA to detect chromosomal abnormalities, including
aneuploidies.
- **Method:** Amplifies DNA with real-time measurement to quantify the number of copies of specific
chromosomes.
- **Application:** Sometimes used in PGT-A to quickly assess chromosomal abnormalities in embryos.

Clinical Applications Of PGT:

Preimplantation Genetic Testing (PGT) has significant clinical applications in reproductive medicine,
particularly for couples undergoing in vitro fertilization (IVF).

1. Reducing the Risk of Genetic Disorders

- **Carrier Couples:** PGT-M is crucial for couples who are carriers of genetic disorders, such as cystic
fibrosis, sickle cell anemia, or Tay-Sachs disease. By screening embryos for these specific genetic
mutations, PGT helps in selecting embryos that are free from the disorder, reducing the risk of the child
inheriting the condition.

2. Preventing Chromosomal Abnormalities

- **Advanced Maternal Age:** Women of advanced maternal age (usually over 35) have a higher risk
of producing embryos with chromosomal abnormalities, such as Down syndrome (trisomy 21). PGT-A is
used to screen for aneuploidies, allowing the selection of embryos with the correct number of
chromosomes, which increases the chances of a healthy pregnancy.
- **Recurrent Pregnancy Loss:** PGT can be applied in cases of recurrent miscarriages where
chromosomal abnormalities might be a contributing factor. By selecting chromosomally normal embryos,
the likelihood of a successful pregnancy is improved.

3. Assisting Couples with Structural Chromosomal Rearrangements

- **Balanced Translocations:** Couples with balanced translocations (where segments of
chromosomes are rearranged but balanced) are at risk of producing embryos with unbalanced
translocations, leading to miscarriage or congenital disabilities. PGT-SR is used to identify and select
embryos with a balanced chromosomal structure, thereby reducing these risks.

4. Improving IVF Success Rates

- **Multiple IVF Failures:** For couples who have experienced multiple failed IVF cycles, PGT-A can
help identify the most viable embryos by ensuring they are chromosomally normal. This can increase the
chances of successful implantation and pregnancy.
- **Single Embryo Transfer (SET):** PGT can support single embryo transfer by selecting the healthiest
embryo, reducing the risk of multiple pregnancies (twins or triplets) while maintaining high success rates

5. Avoiding Sex-Linked Disorders

- **X-Linked Disorders:** Conditions like hemophilia or Duchenne muscular dystrophy, which are
linked to the X chromosome, can be avoided through PGT. PGT-M can be used to screen for these
disorders, ensuring that only embryos without the disease-causing gene are implanted.

6. HLA Matching for Sibling Donor

- **Savior Sibling:** PGT can be used for Human Leukocyte Antigen (HLA) matching to create a “savior
sibling” who can be a compatible donor for a sick sibling needing a stem cell transplant. This involves
selecting an embryo that is both free of the genetic disorder and a suitable HLA match for the existing
child.

7. Sex Selection (Non-Medical)

- **Family Balancing:** In some cases, PGT-A is used for sex selection for non-medical reasons, often
referred to as "family balancing." This is a controversial and ethically debated application, as it involves
selecting embryos based on gender preference.

8. Avoiding Inherited Cancer Syndromes

- **Hereditary Cancer:** Couples with a family history of hereditary cancers, such as BRCA mutations
(linked to breast and ovarian cancer), can use PGT-M to select embryos that do not carry these
mutations, reducing the risk of the child developing cancer later in life.

9. Ensuring Embryo Health in Donor Eggs/Sperm

- **Donor Gametes:** When using donor eggs or sperm, PGT can be employed to ensure that the
embryos created are free from chromosomal abnormalities or specific genetic disorders, providing
additional reassurance to intended parents.

10. Mosaicism Detection

- **Embryo Mosaicism:** PGT, particularly using NGS, can detect mosaicism (where some cells in the
embryo have different genetic compositions). This information can help in decision-making about
whether to implant an embryo with a low level of mosaicism, as these can sometimes still result in a
healthy pregnancy.

Significantlyreduce the risk of genetic disorders and chromosomal abnormalities in their offspring. It
provides valuable information that can improve IVF success rates, prevent miscarriages, and support
informed decision-making in family planning