GENETIC ANALYSIS AND DNA

The continual technological advances and increased knowledge of the human genome puts molecular biology in the forefront for laboratory diagnostics.

DNA testing is a reliable tool for personal recognition and for identifying the presence of bacterial and/or infections. 

DNA ANALYSIS FOR GENETIC PROFILES

PATERNITY TESTS, Consanguinity FOR IMMIGRANTS, etc.: These tests are performed upon appointment

In addition, analyses of rare genetic diseases and infections (e.g. thalassemia, cystic fibrosis, Fragile X, papillomavirus, etc.) can be carried out upon request.

CARDIOVASCULAR RISK EVALUATION

Cardiovascular diseases are one of the most common causes of death in Western countries. However, the most known risk factors such as obesity, smoking, diabetes, high cholesterol and high blood pressure are present in only 50% of the patients afflicted from this disease.

Heredity is an important factor; for many in fact, the main risk factor is simply due to family history of premature cardiovascular diseases. 

The genetic markers of a cardiovascular risk are: 
Factor v fi, Leiden (F 5) Prototrombina (Factor II) and Metilentetraidrofolatoreduttasi (MTHFR) have shown to have a primary role in the onset of these diseases.

Coagulation package/cardiovascular risk

• Factor II (Prothrombin): The activation of prothrombin to thrombin leads to clot formation. There is a genetic variation which is associated to high levels of prothrombin in the plasma leading to an increase of the risk for thrombosis.
• Factor V of Leiden: an essential cofactor which activates Prothrombin to produce Thrombin. 
  The activated Protein C normally inhibits the procoagulant effect, by dividing the activated factor V into three parts. A gene mutation encoding factor V prevents this division.
• MTHER (677C/T and 1298A/C mutations): Methylene tetrahydrofolate reductase (MTHFR) is an enzyme which is involved in the re-methylation of homocysteine to methionine through the intervention of vitamin B12. Rare mutations can cause severe deficiency of MTHFR with the appearance of homocysteine and homocystinuria and low plasma levels of folic acid. This leads to delayed psychomotor development and massive thrombosis. Increased levels of homocysteine in the blood are a risk factor for vascular diseases. Furthermore, in cases of folic acid deficiency in food, the thermolabile variant in MTHFR greatly reduces folic acid in plasma and is therefore a risk factor for neural tube defects in pregnant women.
• PAI-1mutation 4G/5G: Plasminogen Activator Inhibitor 1 (PAI-1) 
  Some studies have shown that the 4G/4G genotype is associated with elevated high plasma levels of PAI-1 which increases the risk of cardio-vascular diseases and preeclampsia in pregnant women.

PRENATAL DIAGNOSIS AND FERTILITY

COUPLES FERTILITY

For Both:

• blood tests: estradiol, FSH, LH, progesterone, prolactin, testosterone, ßHCG.
• Cystic Fibrosis (29 main mutation): occurs mostly within the first years of life. It is a hereditary disease that is transmitted in an autosomal recessive manner, determined by DNA mutations in the gene that encodes the CFTR protein (Cystic Fibrosis Transmembrane Regulator) located on chromosome 7. The genes are inherited in pairs, one resulting from the mother and one by the father. Sick individuals have both pairs of the mutated gene while healthy carriers possess only one copy of the altered gene and one normal. The only way to identify healthy carriers is by performing a DNA test for mutations in the cystic fibrosis gene. The CFTR protein has a fundamental role in regulating the amount of chlorine that is secreted together with biological fluids. In patients with cystic fibrosis, the CFTR protein is either not functional or it is not produced. This means that the secretions of the exocrine glands are more dense and viscous than normal ones, with severe problems for the lungs, digestive system and fertility.
• Chlamydia trachomatis
• Peripheral blood karyotype

For men:

• Y chromosome microdeletions (AZF chromosome): it is responsible for defects in spermatogenesis. A region was located on the long arm of chromosome Y that controls spermatogenesis in males and encodes for the human azoospermia factor AZF.
• Sperm analysis: Identifies number and motility of sperm.
• Spermioculture

FEMALE PREVENTIVE HEALTHCARE MEASURES

• Blood tests: Ca 125, Ca 19.9, ßHCG
• Conventional or ThinPrep PAP smear test. Examination done upon appointment only. 
• Papillomavirus (HPV): Screening
• Papillomavirus (HPV): genotyping (29 strains).

Human Papillomavirus (HPV) infects squamous multilayered epithelia, inducing the synthesis and the production of viral particles inside permissive cells, whose activity is related to the differentiation degree of the host cell.

Around one hundred types of HPV have been identified, more 35 out of these infect the anogenital tract (genital warts), and about 20 are associated with genital cancer. The presence of these sequences is demonstrated in more than 90% of invasive cervical carcinoma cases in squamous cells, in warts and often even in cases of cervical intraepithelial carcinoma.

Based on the pathology (benign or malignant) to which they are most frequently associated, strains of HPV can be divided into:

• low oncogenic risk: HPV 6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81
• high oncogenic risk: HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82.

The most appropriate method for the early detection of cervical cancer is the Pap smear test, which must, however, be combined with the molecular tests which have the purpose of increasing the sensitivity and specificity of the cyto-histological diagnosis, also allowing the typing of papillomavirus. Molecular analysis is the extraction of DNA from a swab/ cytobrush and from the ensuing gene amplification by PCR. In case it is positive, one then proceeds with the genotyping via a specific hybridization to identify which of the above described strains is responsible for the infection.

GYNAECOLOGICAL EXAMS

From vaginal, cervical, urethral and serum swabs

Infections: Qualitative molecular analysis on DNA:

• Papillomavirus (HPV): screening and genotyping (29 strains) if the screening is positive. This exam requires an appointment.
• Chlamydia trachomatis
• Mycoplasma hominis
• Herpes Virus
• Cytomegalovirus
• Toxoplasma

Culture:

• bacteria
• fungi
• Trichomonas
• Gardnerella
• Neisseria gonorrhoeae