The diagnostic applications of molecular biology are extremely varied and, basically, are applicable to any diagnostic problem associated with biological processes. Currently, the main applications are in infectious, oncological, immunological and genetic diseases.
The study of genetic risk factors to multifactorial diseases contributes to the development of early diagnosis tests, new treatments or interventions to avoid the manifestation of the disease or to minimize its severity; in addition, the accurate diagnosis allows the prediction of the risk in the patient’s family which makes the prevention easier.
The predictive or predisposing tests are good to identify the people at risk for a disease before the symptoms appear. These tests are very useful when a person has a family history of a particular disease and there is a method of intervention available to prevent the appearance of the disease or to minimize its severity. The predictive tests are good to identify the mutation that increase a person’s risk to develop a genetic origin disease, as is the case with some types of cancer.
In the GENOMIK Laboratory, we have an increasing list of genetic tests that might be related to infertility process, oncological processes and coagulation disorders. These are fast analysis tests and it only takes a blood sample to determine any factor that modifies the right genetic activity.
Molecular Biology has contributed, not only for the better understanding of infectious diseases, but it has also provided molecular tools for the diagnosis. Through it, it is possible to detect pathogen genomes in biopsy samples or patient fluids. Due to the high affinity of the complementary strands of DNA, it is possible to detect very few organisms in a sample to be analyzed. Two main methodologies are presented in this report: Hybridization with DNA probes and the Polymerase Chain Reaction (PCR). The last one is the most sensitive detection method.
These are some of the PCR technology’s advantages:
Greater sensitivity in the detection of microorganisms difficult to identify by means of conventional methods.
Possibility to characterize with greater discrimination between subtypes of the microorganism of interest.
It provides a quick and reliable diagnosis to establish specific treatments.
Introduction of multiple systems that allow the simultaneous detection by PCR of different infectious agents involved in the same pathology, which reduces costs and execution time.
The range of infectious agents that are detected in GENOMIK, whether bacterial or viral, is wide and will increase according to the needs of the population that we serve and our operational possibilities. The viral load tests are performed through the Real Time PCR technology, which is considered, nowadays, to be very reliable for the quantification of levels of infection by microorganisms such as VIH 1, CMV, Hepatitis B y C, Epstein Barr Virus, Polyomavirus, VPH(16-18-31-33-45-52-56 and 58). Essential information in many cases to define the risk and the therapeutic actions to be undertaken.
The sexual transmission infections represent a serious worldwide public health issue. The World Health Organization estimates that 900 thousand persons are infected every day and 340 million new cases of STI are registered every year in the world, and the proportion of cases is higher among people between 15 to 49 years old, being similar in both genders, with a slight predominance among the men in which they emphasize Syphilis, Gonorrhea, Chlamydia, Human Papilloma Virus and Trichomoniasis.
These infections are generally transmitted from an infected person to another during sexual intercourse without any kind of protection, including vaginal, oral and anal sex. However, some STI are transmitted by other means such as the sharing of syringes in drug addicts, contact with contaminated blood or perinatal transmission from the mother to the child during pregnancy, childbirth or breastfeeding.
Infertility is defined as the inability to complete a pregnancy after a reasonable time of sexual intercourse without taking contraceptive measures; it affects between 15 to 20% of couples of reproductive age. The World Health Organization (WHO), and the European Society of Human Reproduction and Embryology (ESHRE), consider a couple is infertile when the pregnancy does not happen in a two-year minimum term. The conjugal infertility is currently a problem of worldwide distribution with an increasing magnitude for its correlation between the incapacity as such to manage and the psychological factor in the couples who suffer it. Infertility is due to multiple factors that, isolated, may not be important for blocking fertility, but together could be enough to cause reproduction failure.
In Venezuela, 15 out of 100 couples in reproductive age, between 20 and 35 years old, have problems in conceiving a baby. “In 80% of couples who have failed to conceive, there is a proven medical cause”, according to the Gynecology and Human Reproduction doctors.
Currently, multiple causes of infertility are known, such as: immunological mechanisms, genetic alterations and predominantly Sexually Transmitted Infections (STIs), along with changes in sexual behavior (promiscuity), among others.
Therefore, it is important to detect these factors through tests that our personnel is qualified to perform, including:
Multiplex I (MI), where the individual PCR determination of the DNA is performed in a Multiple PCR of 4 main microorganisms causing infertility (Mycoplasma hominis and Ureaplasma urealyticum, Chlamydia trachomatis, Neisseria gonorrhoeae), affecting both men and women. The importance of their determination lies in the similar symptomatology presented by them.
Thrombophilia, we evaluate some genetic mutations that lead to an acquired deficiency being a risk factor for thrombosis during pregnancy and also related to infertility in women. This determination is performed through DNA restriction enzyme cuts and gel detection.
Chromosome Y, allows the determination of different translocations and deletions at the level of Chromosome Y, through a gel detection. These translocations are responsible for the presence of azoospermia (absence of sperm) in the semen, this is directly related to infertility in men. PCR detection of microdeletions in the Y chromosome, one of the main causes of male infertility, is a traumatic event that affects the emotional stability of the individual who suffers it and their marital relationships.
NK cells, n These cells are labeled with a monoclonal antibody directed to their specific membrane antigen and are evaluated through Flow Cytometry. NK cells belong to innate immunity and are found in peripheral blood; however, they are the cells with the highest prevalence in the endometrium. When this cellular population alters its quantity can be the cause of recurrent and spontaneous abortions. This is why it is important in women and their relation to the infertility.
Immunological alterations are those that result from the inadequate behavior of the immune system, a possible functional or structural alteration of genetic origin or any acquired infection. They are capable of causing serious disturbances in the patient, placing them at risk in addition to the disease, making them susceptible to non-habitual or opportunistic infections.
Genomik gives you some of the many tests necessary for an early diagnosis through DNA analysis:
HLA-B27, we make its determination through DNA PCR analysis and gel detection. HLA-B27 is considered a genetic marker for these diseases (spondyloarthropathies).
Leukocyte immunophenotyping (CD3 / CD4 / CD8), for the analysis of the cells we carry out labeling with monoclonal antibodies and are evaluated through Flow Cytometry. These cells are biological indicators of diagnosis and monitoring of some diseases and syndromes, among them: HIV, Tuberculosis, autoimmune diseases and others.
Viral HIV load, using specific sequences directed to the Virus of the Acquired Immunodeficiency, the determination is made through Real Time PCR.
Mainly those aimed at determining paternity, and medical-forensic identification. With more than 10,000 paternity studies performed, GENOMIK Laboratory counts on specialists experienced in this matter and with leading technologies that guarantee the reliability of its results.
We annually participate in international quality control exercises. For this, we have a battery of at least 17 DNA markers, whose use has been validated in the laboratory by means of population studies and altogether offer a superiority to the 99.99% according to the most demanding international standards.
GENOMIK LABORATORY, also offers a range of tests of filial relations such as the Maternity, Brotherhood and Grandmother test. In all of the brotherhood tests, it is necessary the participation of the presumed brothers and it is highly recommended the participation of the progenitor of whom is not in doubt to obtain greater probability of kinship and therefore more reliable results. For example, the mother or the mothers of the alleged brothers (in case these are not common).
Sisterhood and brotherhood tests may be performed without the involvement of the biological parent, however, these may give a lesser probability than the required one to accept it as a conclusive result and in these cases, frequently, we must resort to other genetic markers that provide us more information regarding the maternal or paternal lineage.
We also offer a highly reliable test called the Y chromosome test (Y-STR) to determine if the selected males share the same paternal lineage. The Y chromosome is inherited from parent to child, and this is passed down through generations virtually unchanging Y. The son inherits from the father this chromosome which at the same time will trade it to his son and so on. Therefore, grandfather, son, grandchild and great grandchildren will have the same Y chromosome.
The little or no mutation of the Y chromosome makes this test very accurate. This chromosome is found only in males therefore can only be analyzed to determine the family relationship between males.
In this context, the great challenge of the present moment, in the new century that we have just begun, is to translate into clinical practice the very important advances of our understanding of cancer at the molecular and basic research levels. The task is not easy. On one hand, we know that the molecular and cellular alterations that we have characterized in tumors can affect practically any of the cellular lineages of our organism, producing mutations that normally cannot be avoided. On the other hand, the targets of the malignant mutations are essential genes for the normal growth and development of any cell of the organism. These difficulties suppose theoretical and practical obstacles in designing practical applications of all this knowledge in order to eliminate cancer in our human society. However, new molecular knowledge augurs enormous improvements in the design of new forms of control of tumor diseases. We can predict at least four different levels of approximation of new concrete applications of this knowledge to the control of tumor diseases:
Identification of individuals with genetic predisposition. Due to the genetic load received from their parents, some individuals in the human population have a particularly high risk of tumor diseases. Current knowledge in many cases offers the possibility of significantly reducing the probability of suffering these diseases, through screening procedures that detect the molecular changes that correspond to the initial stages of development of these diseases.
Early and accurate diagnosis of premalignant and malignant changes in patients. The detection of molecular alterations responsible for tumor processes, which in some cases is already technically possible many years before clinical manifestations of them are observed, should produce spectacular benefits in reducing cancer death rates in the coming decades.
In our facilities, we have a line of tests related directly and indirectly with the prognosis of the appearance of genetic alterations that could lead to the appearance of the cancer, as they are:
Molecular diagnosis of HPV and determination of viral load (Causes of cancer of the cervix, the most frequent in the female population of Venezuela).
Taking into account the growing demand in the requests for diagnostic and prognostic tests in the health field, we, at GENOMIK, have opted for the expansion in the variety of molecular biology clinical tests that allow the specialists to give a timely and effective response to their patients in order to be more efficient in the prevention and / or treatments for the control of their pathology.
Soon we will expand our line of work in the oncological area in order to cover the main conditions that are frequent in the Venezuelan population.