LO : Supportive Examination for bacterial infection

Microscopic examination can provide a preliminary, tentative identification of certain infectious agents by revealing their size, shape, and cellular structure. The Gram stain testin which a sample is smeared on a microscopic slide and stained with a special dyeis used to classify all bacteria as either gram positive (blue staining) or gram negative (red staining). The presence of specific inflammatory cells may also provide clues about the type of infection. The acid-fast smear involves spreading a sample on a slide, dyeing it, and treating it with an acid-alcohol solution to identify acid-fast bacteria (which will not be decolorized by the acid-alcohol). This test is often used to examine sputum and support a diagnosis of tuberculosis, or TB (since cultures for TB may take up to 2 months to grow). In addition, microscopic exams are necessary to identify parasites and their eggs. Cultures generally take longer to perform than microscopic examsfrom a day to several weeks but are necessary to identify the organism with certainty and provide a definitive diagnosis. This technique involves placing a specimen in an environment, or medium, that is designed to promote the growth of specific organisms. (The medium is typically a jelly-like substance that contain nutrients to encourage the organisms to reproduce.) Cultures can be used to screen for a wide variety of bacteria, or can be focused to look for specific agents. In some cases, various antimicrobial drugs (such as antibiotics) are then added to a culture to determine which treatment is most effective for killing the offending organism; this is known as drug sensitivity testing.

 Purpose of the Cultures and Microscopic Exams for Infectious Disease To identify the microorganism capable of causing diseases and infections, in some cases, to help determine the best course of treatment The acid-fast bacilli test may be done to monitor treatment for tuberculosis

 Results of Cultures and Microscopic Exams for Infectious Disease The sample is sent to a microbiology or bacteriology laboratory for microscopic examination and/or culturing. In many cases, your doctor will issue a preliminary diagnosis and prescribe a treatment based on your history, symptoms, physical exam, and the results of any initial microscopic studies. Once the final results of a culture and/or drug sensitivity test are in, treatment may be refined accordingly.

 What You Experience during Cultures and Microscopic Exams for Infectious Disease A sample of body fluid, tissue, cells, or stool is obtained and sent to a laboratory for analysis. A variety of procedures may be used, depending on what type of sample is needed. These techniques vary from noninvasivesuch as collecting fluid with a swab (as with a throat culture)to more invasive means involving significant risks, such as tissue biopsy, bronchoscopy, bone marrow aspiration, or lumbar puncture.

Sensitivity test
The goal : The goal of antimicrobial susceptibility testing is to predict the in vivo success or failure of antibiotic therapy. Tests are performed in vitro, and measure the growth response of an isolated organism to a particular drug or drugs. The tests are performed under standardized conditions so that the results are reproducible. The test results should be used to guide antibiotic choice. The results of antimicrobial susceptibility testing should be combined with clinical information and experience when selecting the most appropriate antibiotic for your patient.

The steps
Diffusion disc ATB susceptibility test 1. Prepare bacterial suspension in saline and inoculate it on the surface of Mueller-Hinton agar plate. 2. Apply paper discs with antibiotics according used bacterial species on the inoculated agar using a sterile needle: G+ cocci - penicillin, ampicillin, tetracycline, chloramphenicol, erythromycin, clinda-mycin, G- rods - ampicillin, ticarcillin, tetracycline, chloramphenicol, colistine, gentamicin. 3. Plates are incubated 24 hours at 37oC. 4. In the next exercise you will measure the diameters of zones of growth inhibition. If there is no inhibition zone or the inhibition zone is less than 15 mm in diameter, the tested strain is resistant to the applied antibiotic.

Serologic examination
Serology is a blood test to detect the presence of antibodies against a microorganism. Certain microorganisms stimulate the body to produce antibodies during an active infection.

The steps
Blood is drawn from a vein, usually from the inside of the elbow or the back of the hand. The site is cleaned with germ-killing medicine (antiseptic). The health care provider wraps an elastic band around the upper arm to apply pressure to the area and make the vein swell with blood. Next, the health care provider gently inserts a needle into the vein. The blood collects into an airtight vial or tube attached to the needle. The elastic band is removed from your arm. Once the blood has been collected, the needle is removed, and the puncture site is covered to stop any bleeding.

 In infants or young children, a sharp tool called a lancet may be used to puncture the skin and make it bleed. The blood collects into a small glass tube called a pipette, or onto a slide or test strip. A bandage may be placed over the area if there is any bleeding. The blood is then analyzed in a laboratory to determine how certain antibodies react with specific antigens. The test can be used to confirm the identity of the specific microorganism. There are several serology techniques that can be used depending on the suspected antibodies. Serology techniques include agglutination, precipitation, complement-fixation, fluorescent antibodies, and others.