CECOLFES

Reproductive Molecular
Biogenetics Unit

Patient Information

Information for Patients

Genetics Unit

The genetics service at CECOLFES consists of a modern infrastructure supported by a highly skilled team. It offers highly sensitive and specific state-of-the-art procedures, including the following:  

Cytogenetic Testing: Lymphocyte karyotyping; prenatal diagnosis in amniotic fluid, chorionic villus sampling or chordocentesis; tissue culture of throphoblast or abortion material; genetic analysis in tumor or bone marrow samples, among others.

Biochemical Testing:  Fetal wellbeing and detection of risks for genetic abnormalities using first trimester markers (free beta HCG and PAPP-A) or second trimester triple markers (alpha-fetoprotein, b-fraction chorionic gonadotropin, and non-conjugated striol). Amniotic alpha fetoprotein determination for neural tube defects detection.

Molecular Biology: Chromosomal abnormalities affect male gametogenesis. For this reason, males with abnormal spermograms show Y-chromosome disorders undetectable by conventional lymphocyte karyotyping, the latter being normal in these patients. Using molecular techniques like PCR it is now possible to study the Y chromosome and determine the presence of microdeletions - a cause of infertility in these patients. Other numerical chromosomal abnormalities in chromosomes 13, 18, 21, X and Y, common in patients with abnormal sperm morphology and motility, can also be determined using the FISH technique.

* Cystic fibrosis: Another cause of male infertility is the absence of the vas deferens, and molecular studies have allowed to detect genetic deletions in the cystic fibrosis gene in these patients, 70% of whom have the DF-508 mutation.

* Diagnosis of X-linked diseases: The diagnosis of genetic diseases using molecular techniques has increased since the total sequencing of the human genome. At present it is possible to diagnose all disease for which the responsible genes have been mapped. Using techniques such as PCR it is possible to diagnose genetic abnormalities such as hemophilia A and B, cystic fibrosis, Y-chromosome microdeletions, spinal muscle atrophy, miotonic dystrophy, Rh incompatibility, AZFX / AZFY gene detection for sex selection, sickle-cell anemia. The FISH technique, on the other hand, is used for diagnosing aneuploidies such as X monosomy, trisomy 13, 16, 18 or 21 or Klinefelter syndrome, and structural chromosomal deletions and microdeletions, including the Cri Du Chat, Prader-Willis, Williams, Di-George, Angelman, and Velocardiofacial syndromes. It is also possible to screen for oncogenes such as p53 and c-myc in tumor processes, among others, as well as all abnormalities for which a genetic marker has been described and can be tested using the molecular techniques available in the human genome project.

* Diagnosis of viral and parasitic diseases important during pregnancy using PCR techniques, including toxoplasmosis, cytomegalovirus and rubella.

Pre-fertilization and Pre-implantation Genetic Diagnosis Unit (PGD)

Our PGD program at Cecolfes started in 1995 when we achieved the first birth of a hemophilia-free male in Colombia. Several children free of genetic diseases have been born as part of this program, thus putting an end to the transmission chain of diseases such as hemophilia A and cystic fibrosis. We apply the use of polymerase chain reaction (PCR) and in-situ hybridization (FISH) techniques for diagnosing the couple, followed by pre-implantation genetic diagnosis (PGD) in embryos derived from assisted reproduction techniques (ART). We have ample experience and recognition in this area of pre-fertilization (oocyte and sperm) and pre-implantation (single-cell or blastomere) diagnosis using PCR for genetic abnormalities such as hemophilia A and B, cystic fibrosis, Y-chromosome microdeletions, spinal muscle atrophy, Rh incompatibility, AZFX / AZFY gene detection for sex selection, sickle-cell anemia; or FISH for aneuploidies such as X monosomy, trisomy 13, 16, 18 or 21 or Klinefelter syndrome, and structural chromosomal deletions (translocations). These tests are offered to women over 35 years of age, couples with a history of recurrent miscarriages, spermogram abnormalities, male factor of genetic origin associated with Y-chromosome microdeletions.

Information for Physicians

AMNIOTIC FLUID ALPHA-FETOPROTEIN

Method: Sequential solid phase chemoluminescent immunometric assay

Sample conditions: 3 ml of amniotic fluid, several aliquots.

Sample stability: One week when refrigerated at 4-8°C. One month when frozen.

Days of processing: Daily

Reporting time: 2 days

Reference value: See individual report

Note: The sample must be obtained in patients between 15 and 19 weeks of gestational age.

AMNIOTIC FLUID KARYOTYPING/FISH

Method: Culture in essential fibroblast media.

Sample conditions: Amniotic fluid obtained only at 15-20 weeks.

Sample stability: Up to 72 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 15-20 days

Reference value: See individual report

KARYOTYPING OF OOCYTE REMNANTS OR MISCARRIAGE MATERIAL  

Method: Culture in essential fibroblast media.

Sample conditions: oocyte remnants or miscarriage material.

Sample stability: Up to 72 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 21-28 days

Reference value: See individual report

Note: The sample must be collected in a sterile flask containing sterile normal saline solution (0.9%)

CHORIONIC VILLUS KARYOTYPING

Method: Culture in essential fibroblast media.

Sample conditions: chorionic villi obtained on weeks 10-13.

Sample stability: Up to 24 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 5 days preliminary; 10 days final

Reference value: See individual report

SKIN KARYOTYPING

Method: Culture in essential fibroblast media.

Sample conditions: skin biopsy.

Sample stability: Up to 2-4 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 10-15 days

Reference value: See individual report

Note: The sample must be collected in a sterile flask containing sterile normal saline solution (0.9%)

LYMPHOCYTE KARYOTYPING

Method: Culture in essential lymphocyte media.

Sample conditions: blood with 5-10 ml heparin anticoagulant.

Sample stability: Up to 72 hours

Days of processing: Daily.

Reporting time: 15 days

Reference value: See individual report

Note: Collect a venous sample in green-stopper tube (heparin)

FETAL BLOOD KARYOTYPING (CHORDOCENTESIS)

Method: Culture in essential lymphocyte media.

Sample conditions: blood with 5-10 ml heparin anticoagulant obtained by chordocentesis at 22-25 weeks of gestation.

Sample stability: 24 hours

Days of processing: Daily.

Reporting time: 15 days

Reference value: See individual report

Note: The sample must be collected in a sterile flask containing sterile normal saline solution (0.9%)

BONE MARROW KARYOTYPING

Method: Culture in essential media for metaphase cell isolation.

Sample conditions: bone marrow aspirate with 5 ml heparin anticoagulant.

Sample stability: Send immediately under refrigeration at 4-8°C

Days of processing: Daily.

Reporting time: 15 days

Reference value: See individual report

Note: It is critical to send the sample immediately after it is collected. Please send also peripheral venous blood with heparin anticoagulant.

MITOCHONDRIAL DNA DELETIONS

Method: Polymerase chain reaction (PCR)

Sample conditions: Blood with EDTA anticoagulant (lilac tube), two tubes per patient.

Semen: Total ejaculate in a plastic tube.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 10 days

Reference value: See individual report

Note: Keep refrigerated. Do no freeze. Send in as soon as possible.

LYMPHOCYTE FISH (EITHER CHROMOSOME) FOR DETECTION OF STRUCTURAL OR NUMERICAL ABNORMALITIES.  

Method: Hybridization with specific probes for the target chromosome.

Sample conditions: Blood with 5-10 ml of heparin anticoagulant.

Sample stability: Up to 72 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 5 days

Reference value: See individual report

Note: Collect a venous sample in green stopper tube (heparin)

FISH FOR C-MYC / p53 ONCOGENE

Method: Hybridization with specific probes for the target oncogene.

Sample conditions: Blood with 5-10 ml heparin anticoagulant or tumor tissue in saline solution.

Sample stability: Up to 72 hours when refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 20-30 days

Reference value: See individual report

SPERM FISH (IN-SITU FLUORESCENT HYBRIDIZATION)

Method: Hybridization with probes for chromosomes 13, 18, 21, X and Y.

Sample conditions: Total semen ejaculate in a plastic tube.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 8 days

Reference value: See individual report

Note: Keep refrigerated. Do not freeze. Send in a soon as possible.

FIRST TRIMESTER MARKER

Pregnancy-Associated A Protein, free beta Chorionic Gonadotropin (FREE B-HCG)

Method: Sequential solid phase chemoluminescent immunometric assay.

Sample conditions: 3 ml of serum.

Sample stability: Up to 1 week when refrigerated at 4-8°C. Up to one month when frozen

Days of processing: Daily.

Reporting time: 2 days

Reference value: See individual report

Note: Separate the serum by centrifugation as soon as possible after obtaining the sample. The sample must be collected between 9-12 weeks of gestation. Median values are available for these patients in order to estimate the genetic risk for Down’s Syndrome. Please fill the attached form. Report Nuchal Translucency in order to estimate the risk together with the biochemical markers.

PCR FOR TOXOPLASMA IN AMNIOTIC FLUID

Method: Polymerase chain reaction (PCR) for the detection of the Toxoplasma gondii B1 gene

Sample conditions: Amniotic fluid.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 5 days

Reference value: See individual report

HEMOPHILIA A OR B DIAGNOSIS (CARRIER TESTING)

Method: Polymerase chain reaction (PCR) for indirect detection of intragenetic markers

Sample conditions: Blood with EDTA anticoagulant (lilac tube), two tubes per patient.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 10 days

Reference value: See individual report

Note: Prior consultation is required because of the indirect linkage nature of the method. The number of individuals required varies from one family to another.

CYSTIC FIBROSIS DIAGNOSIS (DELTA F-508 GENE)

Method: Polymerase chain reaction (PCR) for the detection of the Delta F-508 mutation in the CFTR gene (cystic fibrosis transmembrane conductance regulator).

Sample conditions: Blood with EDTA anticoagulant (lilac tube), two tubes per patient.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 10 days

Reference value: See individual report

Note: Keep refrigerated. Do not freeze. Send in a soon as possible.

DIAGNOSIS OF SICKLE CELL ANEMIA

Method: Polymerase chain reaction (PCR) for the detection of the beta globin gene mutation

Sample conditions: Blood with EDTA anticoagulant (lilac tube), two tubes per patient.

Sample stability: Refrigerated at 4-8°C

Days of processing: Daily.

Reporting time: 10 days

Reference value: See individual report

Note: Keep refrigerated. Do not freeze. Send in as soon as possible.

SECOND TRIMESTER TRIPLE MARKER

(Alpha fetoprotein, B-HCG and unconjugated striol)

Method: Sequential solid phase chemoluminescent immunometric assay

Sample conditions: 3 ml of serum

Sample stability: Up to 1 week when refrigerated at 4-8°C . Up to 1 month when frozen.

Days of processing: Daily.

Reporting time: 2 days

Reference value: See individual report

Note: Separate the serum by centrifugation as soon as possible after obtaining the sample. The sample must be collected between 15-19 weeks of gestation. Median values are available for these patients in order to estimate the genetic risk for Down’s Syndrome.

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