1. CONJUCTIVAL DISCHARGE D. M. M. Lab.

2. Conjunctival Discharge Aim of the test An etiological diagnosis of bacterial conjunctivitis by aerobic cultivation with identification and susceptibility test of the isolated bacteria. Types of specimen Two swabs from discharge from the eye(s). Criteria of specimen rejection Inappropriate specimen transport device; mislabeled specimen; unlabeled specimen; specimen received after prolonged delay (usually more than two hour); specimen received in expired transport media.

3. Common pathogensCommensals flora Streptococcus pyogenes Staphylococcus epidermidis Pseudomonas aeruginosaLactobacillus spp Chlamydia trachomatisPropionibacterium spp Streptococcus pneumoniaeStaphylococcus aureus Haemophilus influenzaeVarious Enterobacteriaceae Haemophilus aegypticusVarious streptococcus spp Staphylococcus aureusOccasion pseudomonas aeruginosa Neisseria gonorrheae Streptococcus pyogenes

4. Pre specimen processing Who is authorized to order the test Physician. Quantity of specimen Sufficient amount on swab. Time relapse before processing the sample Eye specimen should be processed immediately because tears contains lysosomes which may kill the organism. Storage Refrigerated (2-8) 0 C.

5. Pre specimen processing Conjunctival discharge Specimen collection I.Pull down the lower eyelid so that the lower conjunctival fornix is exposed. II.Swab the fornix without touching the rim of the eyelid with the sterile cotton swab. III.Place the swab immediately in a bacterial transport medium or, if the specimen is brought to the laboratory immediately, in a sterile test tube with 0.5 mL of phosphate buffered saline.

6. Conjunctival discharge Specimen collection

7. Specimen processing for conjunctiva

8. Direct Visual Examination All material submitted for culture should always be smeared and examined directly by gram stain or other appropriate techniques. Specimen in which chlamydia is suspected can be stained immediately with monoclonal antibody conjugated to fluorescein for detection of elementary bodies or inclusions. Culture Because the constant washing action of the tears the number of organisms recovered from cultures of certain eye infection may be relatively low, so Conjunctival scrapings place directly onto media yield the best results.

9. Specimen processing for conjunctiva One should use blood, MacConkey and chocolate agar plates, because potential pathogen may present in an eye without causing infection it maybe very helpful when any one eye is infected to culture both eyes, If a potential pathogen grows in cultures of both infected and un infected eye the organisms may not be causing the infection, now ever if the organism only grows in culture from the infected eye, it is most likely the causative. Non Culture Methods ELISA and DIFA staining are now available for detection of Chlamydia trachomatis

10. Direct Immuno-fluorescence (DIF) Antibody labeling for Chlamydia trachomatis

11. With Iodine stainWith Giema stain Chlamydia trachomatis Giemsa stain of Chlamydia inclusion bodies (purple "caps" on epithelial cell).

12. Post specimen processing Interfering factors: Patient on antibiotic therapy. Improper sample collection. Result reporting: Report Gram stain finding as an initial report. Report the isolated pathogen and its sensitivity pattern as a final report. Turn around time: Gram stain result should be available half hour after specimen receipt. Isolation of a possible pathogen can be expected after 2-3 days. Negative culture will be reported out 1-2 days after the receipt of the specimen.

13. EAR CULTURE D. M. M. Lab.

14. Ear Culture Aim of the test Etiological diagnosis of otitis externa or otitis media by aerobic and anaerobic culture with identification and susceptibility test of the isolated organism(s). Types of specimen Two swabs from the external or aspiration from middle ear(s). Criteria of specimen rejection Inappropriate specimen transport device; mislabeled specimen; unlabeled specimen; specimen received after prolonged delay (usually more than two hour); specimen received in expired transport media.

15. common pathogens Commensals flora are present in the external ear canal Staphylococcus aureusStaphylococcus epidermidis Streptococcus pyogenesLactobacillus spp. Pseudomonas aeruginosaPropionibacterium spp. Other Gram negative bacilli Staphylococcus aureus Streptococcus pneumoniaeVarious Enterobacteriaceae Haemophilus influenzaeVarious streptococcus spp Anaerobic bacteriaCandida spp. other than albicans Proteus spp.Occasion Pseudomonas aeruginosa

16. Pre specimen processing Who will collect the specimen Medical technologist, Microbiologist for swab from external ear. Otolaryngologist for aspiration from middle ear. Who is authorized to order the test Physician. Time relapse before processing the sample Not more than 2 hours. Storage Refrigerated (2-8) 0 C.

17. Pre specimen processing External Ear Specimen collection 1.Collect a specimen of the discharge on a thin, sterile cotton wool or Dacron swab. 2.Place the swab in a container with the transport medium, breaking off the swab stick to allow the stopper to be replaced tightly. 3.Label the specimen and send it to the laboratory.

18. External Ear Specimen collection

19. Specimen processing for Otitis externa

20. Specimen processing for Otitis media

21. Specimen processing Ear specimen submitted for culture should be inoculated to blood agar, MacConkey, and chocolate agar plates, anaerobic culture also should be setup on those specimens obtained by tympanocentesis or those obtained from patients with chronic otitis media, or mastoiditis. Direct visual examination for material aspirated from the middle ear or mastoid is also examined directly for bacteria or fungi. Cultures from mastoid are generally taken on swabs during surgery and should be transported anaerobically.

22. Additional information For external ear infections only Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa, Vibrio spp. and Aspergillus will be looked for and reported. For middle ear infections only pneumococcus, Streptococcus pyogenes, Haemophilus influenzae and Staphylococcus aureus will be reported with a susceptibility test. For the chronic discharging ear, Bacteroides species and fungi will also be reported in addition to the organisms reported for middle ear infections.

23. Post specimen processing Interfering factors: Patient on antibiotic therapy. Improper sample collection. Result reporting: Report Gram stain finding as an initial report. Report the isolated pathogen and its sensitivity pattern as a final report. Turn around time: Gram stain result should be available half hour after specimen receipt. Isolation of a possible pathogen can be expected after 2-4 days. Negative culture will be reported out 1-2 days after the receipt of the specimen.