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GENERAL PRINCIPLES OF THE  DIAGNOSIS OF INFECTIOUS DISEASES
E tiologic diagnosis of infection  begins with the physical examination of the patient, formulation of the clinical diagnosis and in many cases estimation of the epidemiological circumstances.  Anatomic localization of the infection and the clinical diagnosis suggest variety of possible etiologic agents.  After accurate interpretation of the clinical and epidemiological information, the clinician must select the appropriate specimen, the laboratory tests, and to ensure its adequate transport to the laboratory in order to preserve the viability of the microorganisms.
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Specimen - General Considerations A properly collected specimen is the first important step in the diagnosis of an infection. The results of diagnostic tests depend on the selection, timing, stage of the patients illness, and method of collection of specimen.  Bacteria can be found in different anatomic sites and in different body fluids and tissues during the course of infectious diseases.  Only  sterile  equipment and  aseptic  precaution can be used to avoid contamination of the samples with normal skin flora or saprophytes (e.g.  treatment of the injection site with alcohol  and iodine solutions).
[object Object],[object Object],[object Object],[object Object],[object Object]
A properly collected specimen is very important step in the diagnosis of an infection. The results of diagnostic tests depend upon the selection of specimen, methods of collection, aseptic technique, timing, and transport.  The specimens are collected from different anatomic sites with or without normal bacterial flora.
Methods for identification of infectious agents in clinical specimens ,[object Object],[object Object],[object Object],[object Object],[object Object]
Microscopic Examination  ,[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],Gram  Stain Anthrax  clue cells
Examination of Stained Preparation ,[object Object],[object Object],[object Object],[object Object]
Examination of Stained Preparation ,[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object]
Dark-field microscope   illuminates the object over a dark background and diffracted rays from the specimen make the object visible.  Dark-field microscopy is used for  observation of unstained, live organisms suspended in liquid, which are too thin to be seen by staining   ( T. pallidum, V. cholerae ,  Leptospira   etc . ) .   Leptospira spp. in d ark-field microscopy
[object Object],[object Object],Spores in cells of  B. anthracis
[object Object],[object Object],[object Object],Inclusion bodies of  C. trachomatis  in tissue culture
EM of influenza virus ,[object Object]
Culture Method ,[object Object],[object Object]
Devices for cultivation of microorganisms from clinical specimens Thermostat for aerobic cultivation СО 2  thermostat for cultivation of microaerophils and tissue cultures Jars  for anaerobic and microaerophil atmosphere
Ламинарен бокс  Biohazard  за стерилна работа с тъканни култури и опасни микроорганизми
Identification. Pure Culture and Cultural Characteristics   ,[object Object],[object Object],[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],Biochemical Characteristics
[object Object],[object Object],[object Object]
Instrumentation, Automation and Computer Analysis   ,[object Object],[object Object]
Instrumentation, Automation and Computer Analysis   ,[object Object],[object Object]
A utomated kits and systems  for  diagnostic identification  of microorganisms Mini API Phoenix BD
Immunological Methods for Identification and  Detection of Microorganisms’ Antigens in the Pure   Culture or in the Specimen   ,[object Object],[object Object]
[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],Slide agglutination for determination of salmonella serogroup
Precipitation Tests ,[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object]
Enzyme-Linked Immunosorbent Assay (ELISA) ,[object Object],[object Object]
Enzyme-Linked Immunosorbent Assay (ELISA) ,[object Object]
Direct   Immunofluorescence Test for Antigen Detection   ( inclusion bodies of Chlamidia, isolated in tissue culture )
Direct   Immunofluorescence Test for Antigen Detection ,[object Object],[object Object]
Enzyme-Linked Immunosorbent Assay (ELISA) ,[object Object],[object Object]
Enzyme-Linked Immunosorbent Assay (ELISA) ,[object Object]
ELISA for antigen detection: ,[object Object]
ELISA  for detection of isolated microorganisms or in clinical specimens
[object Object],ELISA  for rotavirus Ag in feces LA for Ag – rotav. in feces, bacteria in CSF IFT – Ag  (Clamidia in specimen from cervix or urethra)
Radioimmunoassay (RIA)  ,[object Object]
Хроматографски тестове за доказване антигените на причинителя в клиничния материал  ( Actim  Influenza A&B :  A   –  H1N1, H3N2  and the avian subtypes  H5N3, H7N3, H9N2 and H5N1  and  B  virus antigen from nasal swab and nasopharyngeal aspirates   in 10 min ) ( също за хламидии, грипни вируси и т.н. ) Step 1 : Addition of  Step 2:  Dipping  Step 3:  Results   extraction buffer
Antimicrobial Susceptibility Testing ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Determination of antibiotic susceptibility of bacteria  D isc-diffusion method
Determination of the minimal inhibitory concentration  ( МIC ) Broth Dilution Method Е-тест
Antibodies Detection  ,[object Object],[object Object],[object Object],[object Object]
Antibodies Detection  ,[object Object],[object Object],[object Object],[object Object]
[object Object]
Indirect Immunofluorescent Test  ,[object Object],[object Object],[object Object]
ELISA  for detection of serum antibodies
Complement-Fixation Test  ,[object Object],[object Object],[object Object]
Western blot  for detection of serum antibodies – confirmation test for HIV, LB etc.
Western blot  in LB Western blot  in HIV
Molecular  M ethods for Detection and Identification of Microorganisms  ,[object Object],[object Object]
[object Object],[object Object],[object Object]
Nucleic Acid Amplification Systems ,[object Object],[object Object],[object Object]
Nucleic Acid Amplification Systems ,[object Object]
[object Object],[object Object]
[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object]

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Diagn.princ.engl. 2011-ok

  • 1. GENERAL PRINCIPLES OF THE DIAGNOSIS OF INFECTIOUS DISEASES
  • 2. E tiologic diagnosis of infection begins with the physical examination of the patient, formulation of the clinical diagnosis and in many cases estimation of the epidemiological circumstances. Anatomic localization of the infection and the clinical diagnosis suggest variety of possible etiologic agents. After accurate interpretation of the clinical and epidemiological information, the clinician must select the appropriate specimen, the laboratory tests, and to ensure its adequate transport to the laboratory in order to preserve the viability of the microorganisms.
  • 3.
  • 4. Specimen - General Considerations A properly collected specimen is the first important step in the diagnosis of an infection. The results of diagnostic tests depend on the selection, timing, stage of the patients illness, and method of collection of specimen. Bacteria can be found in different anatomic sites and in different body fluids and tissues during the course of infectious diseases. Only sterile equipment and aseptic precaution can be used to avoid contamination of the samples with normal skin flora or saprophytes (e.g. treatment of the injection site with alcohol and iodine solutions).
  • 5.
  • 6. A properly collected specimen is very important step in the diagnosis of an infection. The results of diagnostic tests depend upon the selection of specimen, methods of collection, aseptic technique, timing, and transport. The specimens are collected from different anatomic sites with or without normal bacterial flora.
  • 7.
  • 8.
  • 9.
  • 10.
  • 11.
  • 12.
  • 13. Dark-field microscope illuminates the object over a dark background and diffracted rays from the specimen make the object visible. Dark-field microscopy is used for observation of unstained, live organisms suspended in liquid, which are too thin to be seen by staining ( T. pallidum, V. cholerae , Leptospira etc . ) . Leptospira spp. in d ark-field microscopy
  • 14.
  • 15.
  • 16.
  • 17.
  • 18. Devices for cultivation of microorganisms from clinical specimens Thermostat for aerobic cultivation СО 2 thermostat for cultivation of microaerophils and tissue cultures Jars for anaerobic and microaerophil atmosphere
  • 19. Ламинарен бокс Biohazard за стерилна работа с тъканни култури и опасни микроорганизми
  • 20.
  • 21.
  • 22.
  • 23.
  • 24.
  • 25. A utomated kits and systems for diagnostic identification of microorganisms Mini API Phoenix BD
  • 26.
  • 27.
  • 28.
  • 29.
  • 30.
  • 31.
  • 32.
  • 33. Direct Immunofluorescence Test for Antigen Detection ( inclusion bodies of Chlamidia, isolated in tissue culture )
  • 34.
  • 35.
  • 36.
  • 37.
  • 38. ELISA for detection of isolated microorganisms or in clinical specimens
  • 39.
  • 40.
  • 41. Хроматографски тестове за доказване антигените на причинителя в клиничния материал ( Actim Influenza A&B : A – H1N1, H3N2 and the avian subtypes H5N3, H7N3, H9N2 and H5N1 and B virus antigen from nasal swab and nasopharyngeal aspirates in 10 min ) ( също за хламидии, грипни вируси и т.н. ) Step 1 : Addition of Step 2: Dipping Step 3: Results  extraction buffer
  • 42.
  • 43. Determination of antibiotic susceptibility of bacteria D isc-diffusion method
  • 44. Determination of the minimal inhibitory concentration ( МIC ) Broth Dilution Method Е-тест
  • 45.
  • 46.
  • 47.
  • 48.
  • 49. ELISA for detection of serum antibodies
  • 50.
  • 51. Western blot for detection of serum antibodies – confirmation test for HIV, LB etc.
  • 52. Western blot in LB Western blot in HIV
  • 53.
  • 54.
  • 55.
  • 56.
  • 57.
  • 58.
  • 59.