Medical microbiology 2 part
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I GENERAL PRINCIPLES OF MICROBIOLOGICAL DIAGNOSIS
PURPOSE.
To explain the principles of diagnostic medical microbiology and provide an adequate scheme for laboratory diagnosis of infection.
Diagnostic medical microbiology is concerned with the etiologic diagnosis of infection.
1. General principles for specimen collection, transport, storage and processing.
General rules that apply to all specimens for microbiology^
* 71 ie qualitv of the material.must be.adequate, The right sp edmen. the right 1 Lme. the right amount
* The type of specimen to be examined is determined by the presenting clinical picture. The sample should be representative of the infectious process (eg, sputum, not saliva etc.) and-should he taken if possible before antimicrobial drugs are administered.
Contamination of the specimen must be avoided by using sterile equipment and ase11 tic preca ut i ons. Use o f suitab ie steriIe top Is'(d e vie e s) fo r co 11 e ct 1 o n an d -eehnorium^ntahrei-sUfhe^s^qdns-nnisUie label e clmimlmaiEbr.iHdl&d^accQ mparnum by documents (request forms) with the appropriate clinical and other information. The specimen must be taken to the laboratory and examined promptly. Special transport media may7 be helpful and are obligatory for some microorganisms.
Adeequate processing and if necessary storage is of great importance.
Devices for collection: swabs, syringes, blood culture bottles* collection containers for urine, CSF* sputum, washings, aspirates, punctates, tissues etc.
Methods and stages m the,laboratory diagnosis of infection.
Use of appropriate media and conditions for cultivation of microorganisms. Aerobic, anaerobic and microaerophilic cultivation. Incubate the appropriate time,
Inoculation of liquid and solid media. Inoculation methods. Plating out technique for separation of mixed cultures and isolation of pure cultures. Pine culture is an isolated growth of a single, strain free from mixture and contamination with other bacteria'. Each colony constitutes a pure culture, since it consists of the descendants of a single cell, which may be'picked with a sterile inoculating wire (or loop") to prepare a pure a fresh medium.
Jcfentification of microorgatisms
Gram stain or other appropriate stain.
Morphology (shape, size, mutual arrangement), ,
Ability to grow under .aerobic, anaerobic or microaerophilic- conditions.
Growth requirements and characteristic appearance of colonies.
Biochemical properties .
-Identification of species and serovars by use of specific antibodies.
Serological, diagnosis - detection of specific antibodies by agglutination, precipitation. CFT. ELISA and EIA. ASO. immunofluorescence etc.
Antibiotic susceptibility tests. .. .. -
Reporting and interpretation of results
PRACTICAL WORK
1. Tools and devices for collection of specimens for-laboratory diagnosis of infection,
Accompanying documents. Observations, . ■
2. Preparation and staining of smears for microscopic examination, Examinaf :o. A Un-preparations.
Tasks:
Prepare, stain by the staining method of Gram and Loeffler the available microorganisms and draw with the appropriate shape, size, mutual arrangement and colour. If possible guess the microorganism and write it down.
4. Perform., the Gruber agglutination test.
DEMONSTRATIONS AND OBSERVATIONS '
1. Demonstration of different biochemical tests for identification and differentiation of
bacteria. Changed and unchanged test-tubes, demonstrating different biochemical tests. ■-
2. Demonstration of Gruber agglutination test and Widal agglutination test.
3. Demonstration and reading of a disk diffusion test (antibiogram) for susceptibility to antimicrobial agents.' :
4. Demonstration of transport media: Culturette: Falcon tubes. .
Test questions:
1. Which of the following is obligatory in the microbiological diagnosis of:
А/ tuberculosis - Ziehl NeeIsen stain.
В/ bacterial meningitis - Gram stain and Loeffler stain.
С/salmonellosis - Gram stain from stool.
D'AandB -
2. What is characteristic for the Gruber agglutination test? ■
AJ it proves the presence of unknown antigen with the help of known specific antibodies/antibody.
Б/ it proves the presence of unknown antibodies with the help of known homologous antigen.
С/ it is usually performed on a glass-slide.
D/ it is usually performed as part of the microbiological diagnosis of enteric pathogens. B/AandC. ■ ~
F/BandD.
G/ A, C and D.
3. What is characteristic for the Widal agglutination test. ■
AJ it proves the presence.of unknown antigen with the help of brown specific antibodies/antibody.
В/ it proves the presence of unknown antibodies with the help of known homologous antigen.
С/ it is a quantitative reaction.
Di it is usually performed as part of the microbiological diagnosis of tuberculosis.
H/BandC. -
F/B.CandD.
4. Which methods are used for detection of specifis antibodies?
AJ immunofluorescence
В/ co agglutination С/1 atex agglutinati o n D/ none of the above
5. Which methods are used for the detection of specific antigen?
А/ immunobiot
