SUSPICION IN THE FORM OF INFECTION IS THE BASIS FOR SELECTING THE APPROPRIATE METHOD FOR EXAMINING THE TOXOPLASMOSIS DISEASE OF BENDS THAT HAVE NO

Dr. Hiba Riyadh Jameel Al-abodi. Department of Environment, college of sciences, university of Al-Qadisiyah, Iraq. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History Received: 21 July 2018 Final Accepted: 27 August 2018 Published: September 2018


Skin sensitivity Test
This test is known as the delayed skin hypersensitivity test where cellular immunity, which is positive for several months, is used in this test. Toxoplasmin is used as an antigen, injected as a pure solution, and a redness of 5 cm within 48 hours is a positive result (Abd-AL-Hameed, 2007).

Diagnosis by Computerized Tomography Scan (CT.Scan) and Magnetic Resonance Imaging (MRI)
This method is one of the modern methods used to diagnose parasitic infection in the brain, depending on the imaging of the brain tissue, where the diagnosis of any case of inflammation of the brain resulting from the curve of the universe, and found that the use of magnetic imaging MRI is the most sensitive method of CT class. Scan in patients with cerebral palsy (Berroya et al., 2001).

(DT) Sabin Feldman dye test
This type of test is called Methylene blue dye test. This test uses live parasites in its rapid reproductive phase and is the first serological test used to detect the antibodies specific to toxoplasmosis by researchers Feldman and Sabin in 1948 (Tenter et al., 2000). ). The test is easily tested and applied under laboratory conditions. It is used to diagnose toxoplasmosis 1 to 2 weeks after infection and reaches the highest level of antibody after 16-5 weeks. However, it is a disadvantage of this test because it is highly expensive and dangerous to employees because it is used by animals, (Dubey,2007).

Direct heamagglutinatio test
The first to describe and use the DAT is Fulton and Turk in 1959, which is a fast-paced, non-hazardous test using the formalin-infected antigen. Antibodies to the parasite surface are identified as in the most sensitive and specific test of dye (DT) in the discovery of the underlying infection of macular degeneration (Dubey, 2008).

(Modified Agglutination Test (MAT)
Is a test developed and modified by Remington and Desmants in 1980. It was also developed in 1987 by Dubey and Desmant. It is used to detect IgG antibody antibodies by using antigen allergens by destroying IgM antibodies in the patient's serum by using a solution Buffer solution dilute and compound (2-mercapto ethanol)

(IHAT) Indirect Haemagglutination Test
This test was first used (Jacobas and Lunde) in 1957 and is used to kill the infected antigen associated with red blood cells of sheep treated with Pyruvic Aldehyde, as the antibodies bind to sensitive red blood cells.

Latex agglutination test (LAT)
This test is one of the tests to detect antibody antibodies of the conical curved parasite because of its placidity and sensitivity to the host (Salibay et al., 2008). The test kit contains a substance that is attached to the monophyte, soluble molecules as well as parasite antigens. This is because it is used to determine the presence of IgG antibodies as evidence of chronic infection and IgM for latex Antibody Antigen Complex, as well as its ease, sensitivity and privacy, it has the right cost, time and effort to do it (Duck, 2009).

(Complement Fixation Test (CFT)
This test has been widely used in Europe to diagnose toxoplasmosis. Dissolved antigens have been prepared in peritoneal fluid from injected, infected or other animals. However, the defect in this test is the lack of good sensitivity which makes it rarely used today (Rafil , 2008).

Indirect Fluoressnt Antibody Test (IFAT)
This test was used for the first time by Goldman in 1957 to detect conical parasitic antibodies and is widely used but requires a fluorescent microscopy. This test is based on the principle of antigen binding with the specific antibodies found in the patient's serum where it can detect antibodies of the type (IgM), which indicates the presence of acute infection. This test is highly sensitive and easy to administer, allowing positive results at very early stages of the disease, as well as being safer, less expensive and more widely used (Cole et al., 2010).

(Enzyme Linked Immune Sorbent Assay (ELISA)
The first was used in 1971 in the Netherlands by Van-Waeman and Schurs. Nato and Remington have developed the general technique for detecting ELSA antibodies in the blood of the umbilical cord (ELISA, 2004). ELISA includes several types: double ELISA , Micro ELISA, Macro ELISA, and Dot ELISA, which use IgM, IgG, IgG antibody 657 detection is useful for detecting previous infections with elevated IgG antibody in patients with chronic infection. IgM parasite antibodies are the most commonly used for detecting infection When the IgM antibody appears shortly after infection (3-5 days), the ELISA method of IgM does not produce false positive results with a Rheumatoid Factor (RF). Several studies have indicated that the IgM antibody continues for 12 years after infection (Montoya, 2002).
IgE antibodies can be detected in serum ELISA in adults and newborns. Blood IgE concentrations in the blood are lower than those of IgM, IgG. This is useful in the diagnosis of newly acquired infections, and IgA antibody can be detected by ELISA. Remain in the blood for 2-3 months, which is useful in the diagnosis of infection in adults (Wilson et al., 1997).
The antigens are used to treat the antibodies specific to the IgG and IgM antibodies. The IgM antibody is raised shortly after stimulation; decreases more rapidly than the IgG, while the antibody IgG antibody (Yazar et al., 2006). This test was found to be 100% effective, 83.3% (Yazar et al., 2006).

Molecular Tests:
Examples of these tests are Polymerase Chain Reaction (PCR), which has a high ability to diagnose the pathoginc organisms, these tests are highly sensitive as they can diagnose the presence of parasites even if there is one parasite in the sample is also highly susceptible to the dissection of unsatisfactory species of pathogen (Al-abodi et al., 2015). One of the main barriers to using this test is the high cost of these tests, as well as the high level of expertise needed (Sebastiaan et al., 2007). The PCR-based methods have been successful in detecting the three species of lamp and have been selected for clinical studies (Calderaro et al., 2006 ; Al-Mayali and Al-abodi, 2017). This technique has evolved into a number of methods that are derived and the most recent and accurate is the polymerase reaction technique in real time. The following is a definition of this technique:

Real time polymerase chain reaction (RT_ PCR)
A new scientific technique combining the amplification of the DNA sample with the detection of the amount and quality of the amplification output in a single reaction tube, which protects the samples from contamination by opening and closing the reaction tubes ((Abdul-Ghani, 2011). This technique is based on double dinucleotide reagents, which are a radioactive dye associated with each detector that has a sequence complementary to one of the resulting tapes. A fluorescent dye, called a Reporter, is connected to the 5-pin reagent the third part of the reagent is connected to another dye called the Quencher dye. When the laser is emitted by laser or halogen, the dye absorbs the energy of the photons and then transfers it to the absorbent dye via the Fluoresce resonance energy transfer (FRET). (Shibli et al., 2008) Basics of polymerase chain reaction technology in Real-Time PCR: The RT-PCR method was first used by Higuchi and his group (1993). The RNA is usually the basis of the real time reverse transcriptase technique and cannot be used as a template in RT-PCR. Therefore, the first step is to transform the template RNA to cDNA in the reverse transcription process through which the gene expression can be measured by RT-PCR technology that can control real-time DNA amplification. This process is carried out by means of special chemicals and tools consisting of a special radioactive dye and sensors that include:

Green Cyberspace Syber green I dye:
It is the most common pigment in the RT-PCR device and is associated with a non-specific type of DNA tape (Abdul-Ghani, 2011; Menotti et al., 2010), which is non-radioactive when placed in a normal solution but lighted and increased 1000 fold when connected with the dsDNA The green cypress dye is also associated with the polymerase chain reaction products that lead to increased radioactive dye in sequence and can then be determined.
658 Figure 1:-Cyber green dye is increasingly radioactive when its parts are connected with the dsDNA tape (Dorak, 2006).

Hydrolysis probe dyes:
The TaqMan

Hybridization Technique:
The probe senses the Flourochrome dye at the third end and the other senses a second dye when the two colors are in a similar space (approximately 0-1 nucleotides) and that the light emitted from the dyed dye will evoke the receiving dye and result in the emission of the fluorescent dye can be determined during the annealing phase. In the first part of the fermentation phase of polymerase chain reaction and after each successive cycle of PCR, the number of hybrid and polycarbonate probes will increase and result in more brilliance. Joint processes between reverse transcription Real-time polymerase chain reaction is very effective when gene expression is measured (Dorak and Tervli, 2009). to analyze the results using RT-PCR, two methods are used: First: absolute quantity. Threshold Cycle (Ct) tells the first signal or radiation of the dye when it binds to nucleic acids when DNA is doubled. This cycle is measured at the exponential phase, for the amplification zone the PCR output is multiplied by each cycle according to Figure (4). There are many applications for the use of RT-PCR in medical fields, for example to diagnose HIV (Human Immunodeficiency Virus) (Menotti et al., 2010) and in laboratories for the diagnosis of incurable diseases such as avian and swine flu.