PULGUIFERTI Laboratory PDF

# Laboratorio Pulguiferti ## Introduction The Pulguiferti Laboratory will carry out embryonic tests before birth, detecting genetic diseases that the embryo may have. These diseases can have serious consequences for the baby. Pregnant women need to receive adequate prenatal care and undergo regula...

# Laboratorio Pulguiferti ## Introduction The Pulguiferti Laboratory will carry out embryonic tests before birth, detecting genetic diseases that the embryo may have. These diseases can have serious consequences for the baby. Pregnant women need to receive adequate prenatal care and undergo regular examinations to detect possible problems early on. ## Prenatal testing These are blood tests performed at the beginning of pregnancy to look for diseases or infections that could affect the health of the mother and the baby. Results can help guide treatments to prevent serious complications. The most comprehensive tests are blood-based DNA tests. ## Invasive techniques These techniques involve applying micromanipulation to the embryo. A surgical procedure is performed through an external object, such as a needle, a probe, a device, or an endoscope. It's worth noting that these techniques pose a small risk of miscarriage or infection. ### Amniocentesis This involves a transabdominal puncture between weeks 15 and 20 of gestation, extracting amniotic fluid from the amniotic sac to obtain the fetal karyotype. ### Chorionic villus sampling Chorionic villus sampling is carried out between weeks 11 and 13 of gestation. Using ultrasound guidance, a segment of chorionic villus tissue from the developing placenta is extracted. This helps determine if the fetus has chromosomal abnormalities, but it doesn't detect other congenital anomalies. * **Transcervical** is performed through the cervix, using a flexible plastic catheter or a malleable metal forceps. * **Transabdominal** involves inserting a 17/19G or 18/21G trocar through the abdomen, obtaining samples from different parts of the placenta. ### Cordocentesis Cordocentesis is a procedure that obtains fetal blood through a puncture in the umbilical cord vessels, guided by ultrasound or fetoscopy. It is performed from week 20 of gestation onwards, allowing for rapid analysis and studies on the fetus's chromosomes, immune system, as well as specific hemopathies. ### Fetal tissue biopsy A guided ultrasound puncture is used on an anesthetized mother. It is rarely performed, only when specific genetic studies are required for a specific organ. ### Embriofetoscopy This involves direct observation of the fetus through an endoscope. It’s not often performed because it’s risky for both the mother and the fetus, but it can be utilized to perform a biopsy on the fetal tissues. ## Non-invasive Techniques These techniques pose no risk to the mother or the fetus because they do not invade the body. They detect chromosomal abnormalities in the fetus by analyzing small DNA fragments found in the mother’s blood. ### Fetal DNA Test in maternal blood This test, performed from weeks 9 or 10 of gestation, involves drawing a sample of the mother's blood to detect fetal DNA and possible genetic abnormalities. ### Combined first-trimester screening Known as Triple Screening, it involves a combination of ultrasounds and blood tests (PAPP-A and beta-hCG) . It analyzes markers to assess the risk of chromosomal abnormalities in the fetus. ## Embryonic diseases The document presents a table with several syndromes and their explanations. ### Tetralogy of Fallot A heart condition associated with a microdeletion and duplication on chromosomes 22 and 11. ### Dextrocardia A non-syndromic effect on chromosome 14, characterized by the heart's location in the right chest cavity. ### Achondroplasia A mutation in the fibroblast growth factor receptor 3 (FGFR3) gene, affecting bone growth, particularly in the long bones. ### Down Syndrome Trisomy 21, caused by an extra copy of chromosome 21. ### Huntington's disease An autosomal dominant disorder affecting chromosome 4, caused by repeating CAG sequences in DNA. ### Fragile X syndrome A genetic condition affecting the FMR1 gene on the X chromosome, predominantly affecting males. It’s linked to physical and intellectual challenges, especially in older men. ### Klinefelter syndrome Caused by an extra X chromosome, present in males (47 XXY) affecting physical development, and reproductive system. ### Cleft lip and palate A genetic variation on chromosome 8, affecting the development of the lip and palate, possibly influenced by environmental factors. ### Hemophilia A genetic condition affecting the blood coagulation process, transmitted through the X chromosome. ### Muscular dystrophy A group of genetic disorders affecting muscles, typically caused by changes in the DNA sequence responsible for muscles' protein. ### Patau Syndrome Trisomy 13, characterized by an extra copy of chromosome 13. ### Edwards Syndrome Trisomy 18, characterized by an extra copy of chromosome 18. ## Technical procedures performed in the lab This section lists the techniques performed in the lab and explains them in detail. ### FISH technique Fluorescence in situ hybridization (FISH) is a method for marking chromosomes with probes that emit fluorescence. These probes allow for the visualization, differentiation, testing of chromosomes in metaphase or interphase, and analysis of possible anomalies. ### PGD (Preimplantation Genetic Diagnosis) PGD is a technique implemented in the chromosomes of parents, detecting the presence or absence of specific areas in each embryo, identifying genetic abnormalities that cause diseases, such as Down Syndrome, Turner Syndrome, Klinefelter Syndrome, or single-gene diseases like Fragile X syndrome, spinal muscular atrophy, cystic fibrosis, phenylketonuria, and albinism. ### CGH-array This technique is used for detecting chromosomal imbalances that are too small to be detected with the karyotype. ### PCR (Polymerase Chain Reaction) The PCR technique copies a specific DNA segment through cycles of heating and cooling, creating millions of copies in an hour. If viruses or other pathogens are present, the equipment can detect them. ### Karyotype This test determines the size, shape, and number of chromosomes in a cell. It involves examining a sample of cells from the body and provides information about the individual's genome. ## Work areas ### Laboratory * The lab is the area where samples are organized, classified, and prepared for analysis. If necessary, samples are sent to other laboratories. * The reception area is where samples are received, checked against the order, and the quality of the samples is verified. Samples not meeting the requirements are rejected. * The pre-treatment area is where samples are prepared before full processing. ### Pre-PCR area This is the “dirty” lab for working with DNA, where all the reaction reagents are prepared for PCR. ### Personnel Washing/Sterilization Area These are controlled areas designed to prevent microbial contamination during the handling of pharmaceutical, medical, and biological products. ### PCR and post-PCR area This area is dedicated to manipulating samples after PCR is complete. The main goal is to prevent cross-contamination and ensure the integrity of the results. ### Darkroom This is a darkened room where light-sensitive materials such as photographic films and paper are processed. It’s also where FISH techniques, which utilize fluorescence, or a laser microscope are performed. ### Culture room This is an isolated room with climate-controlled airflow filtered by HEPA, creating a positive air pressure. This prevents contaminants entering the room, maintaining a clean environment for cell cultures used to obtain karyotypes. ### Changing room This area is provided for employees to change, storing their workday or personal clothes. ### Bathrooms This room is for personal hygiene, showering and disposing of waste. ### Reception area This acts as the secretariat, generally located between the waiting area and the sampling area. In smaller laboratories, it serves as the entrance with direct access from the exterior. ### Storage area * **Ambient temperature area:** This area stores disposable materials and reagents that do not require special storage conditions. * **Cold rooms:** These are maintained between 2°C and 8°C, designed to store reagents and samples that require refrigeration. * **Freezers:** These are used for storing samples that require freezing. The document presents a diagram of the laboratory layout. ## Equipment * **Thermal cycler:** This device is used to automate and execute temperature cycles for DNA amplification in PCR, encompassing the steps of denaturation, hybridization, and extension. * **Fluorescence microscope with imaging capture and analysis systems:** This microscope uses a specific wavelength of light to excite fluorescently labeled substances, producing images of higher resolution and contrast. It incorporates a capture system to record images or videos and save them to a computer. * **Centrifuge:** This machine separates substances based on their density through centrifugal force, commonly used in clinical laboratories. * **Laminar flow hood:** This device protects the laboratory personnel, materials, and the environment from biological contaminants. It features a filter that directs a constant, unidirectional flow of clean air, reducing contamination risk. ## Materials * **Petri dishes:** These circular containers with lids are used for culturing bacteria, fungi, and other microorganisms. * **Eppendorf tubes:** These small tubes are ideal for storing small volumes of liquids, primarily for PCR procedures. * **Pipette with suction device:** This tool is used to transfer accurate volumes of liquids with controlled suction, generally used for transferring liquids between laboratory containers. * **Micropipette:** This device is designed for precise and automatic transfer of very small volumes of liquids, primarily used for sample collection and dilution, utilizing disposable tips for increased precision. * **Pipette tips:** These are disposable tips used with micropipettes, promoting sterile procedures and preventing cross-contamination. * **Slides and coverslips:** These are small glass plates used to prepare samples for viewing under a microscope. Slides are used as a base, while coverslips cover the sample on top. ### Other Materials * **PCR plates, strips, and films:** These are designed for PCR and designed to minimize sample evaporation, ensuring accurate analysis, especially with small volumes. * **Sterilization controls:** These help assess the effectiveness of the sterilization processes used in a lab. * **Personal Protective Equipment (PPE):** PPE, such as gloves, glasses, masks, and lab coats, ensures protection from external hazards, including viruses, bacteria, gases, and other substances. ## Additional Content The document also includes several sections: * **Interesting facts and videos** * **Bibliography** * **Techniques and procedures** * **Work areas** * **Equipment and materials** The document also provides a comprehensive list of online resources about the topics discussed. This information is presented with a focus on clarity and structure. The aim is to offer a comprehensive and practical guide to the Laboratory's operation and procedures.