Use of Sputum sample for diagnosis of disease, interpretation, treatment & clinical corelation
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Sputum Sample Sputum is mucus that is coughed up from the lower airways.In medicine, sputum samples are usually used for microbiological investigations of respiratory infections and cytological investigation of respiratory systems.
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Use of Sputum sample for diagnosis of disease, interpretation, treatment & clinical corelation
- 3. What is Sputum ? • Sputum is mucus that is coughed up from the lower airways. • (Phlegm, mucus) Sputum definition - Medical Dictionary definitions of popular medical terms easily defined on MedTerms
- 4. Types Of Sputum • Bloody • Rusty colored • Purulent • Foamy white • Frothy pink Sputum Color is the Key to Treating Acute COPD Exacerbations
- 6. Diagnosis of disease using Sputum • Smear ----Gram Stain----Microscopy • Culture -----Microscopy • Others (PCR, etc.)
- 9. The specimen that is uniformly green and purulent presents no problem. Any portion of it will provide useful information.
- 10. These are specimens that consists mainly of saliva with a small amount of material that may be sputum. The flecks should be selected.
- 11. Specimens may be heavily stained with blood. Select the portion of the sample that is mucoid and blood stained.
- 12. When only a few blood stained flecks are present, these are the portions to select.
- 13. Avoid any part of the specimen that is watery, as this is probably saliva.
- 14. Collection of Sample Use a sterile cotton swab to extract the specimen from the container.
- 15. 1 2 4 5 3 6 7 8 9
- 16. Microscopy
- 17. Possible Infections • Bacterial(pneumonia, whooping cough, Legionnaire's disease, an atypical pneumonia, parrot fever, TB, otitis media, bronchitis, sinusitis, laryngitis ) • Fungal (Aspergillosis, Blastomycosis, Coccidioidomycosis, Cryptococcus, Histoplasmosis ) • Viral (In combination with certain bacteria)
- 18. Possible Bacterial Pathogens • Streptococcus pneumoniae • Staphylococci • Hemophilus influenzae • Members of the family Enterobacteriaceae • Some pseudomonads • Moraxella catarrhalis • Mycobacterium tuberculosis
- 19. Mucus
- 22. Streptococcus pneumoniae • cause of bacterial pneumonia • organisms are arranged in pairs and are lancet shaped • surrounded by a large capsule • organisms are reported as Gram positive cocci in pairs.
- 24. Staphylococcus aureus • They are usually round or slightly oval cocci which occur singly in small clusters. • Report these organisms as Gram positive cocci in pairs and clusters.
- 27. Haemophilus influenzae • The presence of many very tiny pleomorphic Gram negative rods is strongly suggestive of Hemophilus influenzae. • They should be reported as small, pleomorphic Gram negative rods.
- 30. Enterobacteriaceae • These organisms are gram negative rods • They are usually fat rods, larger than Hemophilus sp.
- 33. Pseudomonas sp. • Pseudomonads are usually long slender Gram negative rods. • Like Enterobacteriaceae, their morphology is not sufficiently typical to be able to characterize them on a Gram smear.
- 34. • Gram-negative, aerobic, oxidase-positive diplococcus
- 35. Moraxella catarrhalis • Gram Negative, Rod shaped • Diplococcus
- 38. Mycobacterium Tuberclosis •M. tuberculosis is characterized by granulomas containing Langhans giant cells, which have a "horseshoe" pattern of nuclei. •Organisms are identified by their red color on acid-fast staining.
- 40. Yeast • Yeasts and pseudohyphal elements are frequently seen. • Small numbers. • Increased numbers are often seen in patients who are immuno-logically compromised and in those who have been extensively treated with antibiotics. • These patients often have oral candidiasis. • Yeasts are much larger than bacteria. Notice the size difference between the yeast and the gram negative rods.
- 41. Viral Infection • Viruses are a common cause of pneumonia. For a viral culture, sputum is mixed with commercially-prepared animal cells in a test tube. Characteristic changes to the cells caused by the growing virus help identify the virus. The time to complete a viral culture varies with the type of virus. It may take from several days to several weeks.
- 42. Media Culture used for sputum • Mac Conkey Agar • Chocolate Agar • Blood Agar
- 43. Clinical Correlation • Moraxella catarrhalis - : otitis media, bronchitis, sinusitis, laryngitis.
- 44. Streptococcus pneumoniae- • acute sinusitis, otitis media, conjunctivitis, meningitis, bacteremia, sepsis, osteomy elitis, septic arthritis, endocarditis, peritonitis,pericarditis, cellulitis, and brain abscess.
- 45. Mycobacterium tuberculosis • (Mycobacterium africanum,Mycobacterium bovis (Dassie's bacillus), Mycobacterium caprae, Mycobacterium microti, Mycobacterium mungi, Mycobacterium orygis and Mycobacterium pinnipedii) – TB
- 46. Haemophilus influenzae - • (oppurtunistic pathogen) bacteremia, pneumonia, epiglottitis and acute bacterial meningitis, cellulitis, osteomyelitis, and infectious arthritis.
- 47. Fungal Diseases • Aspergillosis, Blastomycosis, Coccidioidomycosis, Cryptococcus, Histoplasmosis
- 48. Treatment • Antibiotics (cefotaxime , ceftriaxone ampicillin , sulbactam, cephalosporins , rifampicin, izoniacid,etc.) • Vaccines (BCG, Hib conjugate vaccine , etc. )
- 49. Mucus • "Mucus is incredibly important for our bodies, It is the oil in the engine. Without mucus, the engine seizes." Explains Michael M. Johns, III, MD, director of the Emory Voice Center and assistant professor of otolaryngology -- head and neck surgery at Emory University.
- 50. References • Sputum definition - Medical Dictionary definitions of popular medical terms easily defined on MedTerms • Clinical Microbiology procedures handbook, American Society for Microbiology 2nd Ed. 2007 update • Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc. ISBN 0-07-140923-8. • Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc. ISBN 0-07-140923-8. • Sputum Color is the Key to Treating Acute COPD Exacerbations • Mawas F, Ho MM, Corbel MJ (January 2009). "Current progress with Moraxella catarrhalis antigens as vaccine candidates". Expert Rev Vaccines 8 (1): 77–90. doi:10.1586/14760584.8.1.77.PMID 19093775. • Yu S, Gu XX (June 2007). "Biological and Immunological Characteristics of Lipooligosaccharide-Based Conjugate Vaccines for Serotype C Moraxella catarrhalis". Infect. Immun. 75 (6): 2974–80.doi:10.1128/IAI.01915- 06. PMC 1932890. PMID 17371852 • Winstanley, T.G. and R.C. Spencer. “Moraxea catarrkar: antibiotic susceptibility with special reference to trimethoprim.” Journal of Antimicrobial Chemotherapy 18, no. 3 (1986): 425-426. • Helminen, Merja E., Isobel Maciver, Jo L. Latimer, Julia Klesney-Tait, Leslie D. Cope, Maria Paris, George H. McCracken, Jr., and Eric J. Hansen. “A Large, Antigenically Conserved Protein on the Surface of Moraxella catarrhalis Is a Target for Protective Antibodies.” The Journal of Infectious Diseases 170, no. 4 (1994): 867-872. • h Melendez, Philipp R. and Royce H. Johnson. “Bacteremia and Septic Arthritis Caused by Moraxella catarrhalis.” Reviews of Infectious Diseases 13, no. 3 (1991): 428-429. • Maciver, Isobel, Manaswat Unhanand, George H. McCracken Jr., and Eric J. Hansen. “Effect of Immunization on Pulmonary Clearance of Moraxella Catarrhalis in an Animal Model.” The Journal of Infectious Diseases 168, no. 2 (1993): 469-472.
- 51. • Kuhnert P; Christensen H (editors). (2008).Pasteurellaceae: Biology, Genomics and Molecular Aspects. Caister Academic Press. ISBN 978-1-904455-34-9. • Ryan KJ; Ray CG (editors) (2004). Sherris Medical Microbiology (4th ed.). McGraw Hill. pp. 396–401. ISBN 0-8385-8529-9. • Slack, MP; et al. (1998). "Enhanced surveillance of invasive Haemophilus influenzae disease in England, 1990 to 1996: impact of conjugate vaccines.". Pediatr Infect Dis J. 17 (9 suppl): S204–7. PMID 9781764. • Behrman, Richard E.; Robert M Kliegman and Hal B. Jenson (2004). Nelson Tratado de Pediatría. Elsevier. p. 904. ISBN 84- 8174-747-5. Retrieved 2009-09-11. • Puri J, Talwar V, Juneja M, Agarwal KN, Gupta HC (1999). "Prevalence of antimicrobial resistance among respiratory isolates of Haemophilus influenzae". Indian Pediatr 36 (10): 1029–32. PMID 10745313. • Levine OS, Schuchat A, Schwartz B, Wenger JD, Elliott J; Centers for Disease Control (1997). "Generic protocol for population-based surveillance of Haemophilus influenzae type B". World Health Organization. WHO/VRD/GEN/95.05. • John TJ, Cherian T, Steinhoff MC, Simoes EA, John M (1991). "Etiology of acute respiratory infections in children in tropical southern India". Rev Infect Dis 13: Suppl 6:S463–9.PMID 1862277. • Kennedy WA, Chang SJ, Purdy K, LE T, Kilgore PE, Kim JS et al. (2007). "Incidence of bacterial meningitis in Asia using enhanced CSF testing: polymerase chain reaction, latex agglutination and culture". Epidemiol Infect 135 (7): 1217– 26. doi:10.1017/S0950268806007734.PMC 2870670. PMID 17274856. • Pericone, Christopher D., Overweg, Karin, Hermans, Peter W. M., Weiser, Jeffrey N. (2000). "Inhibitory and Bactericidal Effects of Hydrogen Peroxide Production by Streptococcus pneumoniae on Other Inhabitants of the Upper Respiratory Tract". Infect Immun 68 (7): 3990–3997.doi:10.1128/IAI.68.7.3990-3997.2000. PMC 101678.PMID 10858213. • Lysenko E, Ratner A, Nelson A, Weiser J (2005). "The role of innate immune responses in the outcome of interspecies competition for colonization of mucosal surfaces". PLoS Pathog 1 (1): e1. doi:10.1371/journal.ppat.0010001.PMC 1238736. PMID 16201010. • Chang CM, Lauderdale TL, Lee HC, et al. (August 2010)."Colonisation of fluoroquinolone-resistant Haemophilus influenzae among nursing home residents in southern Taiwan". J. Hosp. Infect. 75 (4): 304– 8.doi:10.1016/j.jhin.2009.12.020. PMID 20356651.
Editor's Notes
- Phlegm is in essence a water-based gel consisting of glycoproteins, immunoglobulins, lipids and other substances. Important part of the normal defense mechanism of the lungsProduced by goblet cells which line the bronchiSpreads in a thin, even layer over the tissue surfacesProvides a transport system for the removal of foreign bodies from the respiratory tractIn many disease states, an excess of mucus is produced and is expectoratedCells and bacteria become enmeshed in the mucus and it is this combination of material that is known as sputumSputum:One of the most common types of specimen submitted to the laboratory for bacterial examination. Difficult to obtain because of contamination with saliva.Even specimens collected by bronchoscopy or through an endotracheal tube may be mixed with oropharyngeal secretions.Many of the bacteria which are known to cause lower respiratory tract infections may be present in the oropharynx as part of the normal flora e.g., staphylococci, pneumococci, and gram negative rods. The examination of a direct smear from the specimen can be very helpful in diagnosing respiratory infections and in determining the usefulness of the information provided by the culture.
- . The colour can provide hints as to effective treatment in Chronic Bronchitis Patients:
- Equipment requiredThe equipment needed to prepare the smears consists of two slides (preferably ones with frosted ends) a package of sterile swabs, a pencil a Bunsen burner. Label the two slides with the patient’s name, the date of collection, and the specimen accession number.
- Specimens may be primarily clear and slightly viscid with flecks of white or greenish material embedded. These are specimens that consists mainly of saliva with a small amount of material that may be sputum. The flecks should be selected.
- The sample to be used may be drawn up the side of the container with the swab to make selection easier. Mucus can be "cut" with the swab by drawing the swab against the side of the container and thus separating off part of the mucus. Merely dipping the swab into the specimen will not usually provide the best sample.
- Fix the smear by passing it through a Bunsen flame several times. Do not allow the smear to be in contact with the flame for more than one second at a time. 5 or 6 passes through the flame is sufficient. The slide should be warm, not hot, to the touch.Stain one of the smears using the Gram stain procedure. Sputum smears are usually thicker than most other types of smear and often need more decolorization than usual. Decolorize until all or most of the blue color is removed
- Mucus is easily seen under low power magnification. It forms long irregularly shaped strands which may have cells and bacteria embedded in it.
- The etiology of bacterial infections cannot be diagnosed solely by the Gram stain morphology of the organisms in a direct smear. However, the morphology of some organisms is sufficiently characteristic that a tentative diagnosis can be made on the basis of the smear .
- Mucus is easily seen under low power magnification. It forms long irregularly shaped strands which may have cells and bacteria embedded in it.
- Hemophilusinfluenzaeoften stains very weakly and tends to blend into the background material in the smear and it is easy to overlook them. On this slide, they are most clearly seen as intracellular organisms in the poly with a 4-lobed nucleus. Look for more amongst the background material. Pleomorphism (cytology): variability in the size and shape of cells and/or their nuclei
- Clinical Significance : These bacteria are known to cause otitis media,[2][3] bronchitis, sinusitis, and laryngitis. Elderly patients and long-term heavy smokers with chronic pulmonary disease should be aware that M. catarrhalis is associated with bronchopneumonia, as well as exacerbations of existing chronic obstructive pulmonary disease (COPD).It has also been known to cause infective exacerbations in adults with chronic lung disease, and it is an important cause in acute sinusitis, maxillary sinusitis, bacteremia, meningitis, conjunctivitis, acute purulent irritation ofchronic bronchitis, urethritis, septicemia (although this is rare), septic arthritis (which is also a rare occurrence), as well as acute laryngitis in adults and acute otitis media in children.[6][7] M. catarrhalis is an opportunistic pulmonary invader, and causes harm especially in patients who have compromised immune systems or any underlying chronic disease
- Phlegm is in essence a water-based gel consisting of glycoproteins, immunoglobulins, lipids and other substances. Important part of the normal defense mechanism of the lungsProduced by goblet cells which line the bronchiSpreads in a thin, even layer over the tissue surfacesProvides a transport system for the removal of foreign bodies from the respiratory tractIn many disease states, an excess of mucus is produced and is expectoratedCells and bacteria become enmeshed in the mucus and it is this combination of material that is known as sputumSputum:One of the most common types of specimen submitted to the laboratory for bacterial examination. Difficult to obtain because of contamination with saliva.Even specimens collected by bronchoscopy or through an endotracheal tube may be mixed with oropharyngeal secretions.Many of the bacteria which are known to cause lower respiratory tract infections may be present in the oropharynx as part of the normal flora e.g., staphylococci, pneumococci, and gram negative rods. The examination of a direct smear from the specimen can be very helpful in diagnosing respiratory infections and in determining the usefulness of the information provided by the culture.