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Sputum Culture: D. M. M. Lab

The document discusses the process for performing a sputum culture to diagnose lower respiratory tract infections, including collecting and processing sputum samples, examining samples under a microscope, culturing samples to identify bacteria, and identifying common pathogenic bacteria found in sputum such as Streptococcus pneumoniae and Haemophilus influenzae. The quality of sputum samples is important, and samples should minimize contamination from the oropharynx while containing sputum, pus, or blood for accurate diagnosis.

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665 views28 pages

Sputum Culture: D. M. M. Lab

The document discusses the process for performing a sputum culture to diagnose lower respiratory tract infections, including collecting and processing sputum samples, examining samples under a microscope, culturing samples to identify bacteria, and identifying common pathogenic bacteria found in sputum such as Streptococcus pneumoniae and Haemophilus influenzae. The quality of sputum samples is important, and samples should minimize contamination from the oropharynx while containing sputum, pus, or blood for accurate diagnosis.

Uploaded by

Subha
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPT, PDF, TXT or read online on Scribd
  • Introduction to Sputum Culture: Introduces the purpose of sputum culture in diagnosing respiratory infections.
  • Types of Specimen: Describes different types of sputum and characteristics used for diagnostic purposes.
  • Specimen Collection and Handling: Details methods for collecting sputum specimens and criteria for rejection to ensure quality samples.
  • Processing Methods: Explains the methodology for processing sputum specimens including staining techniques.
  • Staining Techniques: Covers techniques and principles of staining for identifying specific bacteria in sputum.
  • Result Interpretation and Reporting: Explains how to interpret and report results from sputum cultures, including turnaround times.

SPUTUM CULTURE

D. M. M. Lab.
Sputum Culture
An etiological diagnosis of lower respiratory tract infection by microscopic
examination and culture with identification and susceptibility test of the
isolated organism.
Sputum is a thick fluid produced in the lungs and in the airways leading to the
lungs.

Sputum can be:


I. Bloody (often found in tuberculosis) (Hemoptysis).
II. Rusty colored - usually caused by pneumococcal bacteria (in pneumonia).
III. Purulent - containing pus:
a. a yellow-greenish (mucopurulent) color.
b. a white, milky, or opaque (mucoid).

IV. Foamy white - may come from obstruction or even Edema.


Types of specimen
 Expectorated, Transtracheal aspirates, Translaryngeal aspirates, Bronchoalveolar
Lavage.
The common bacterial pathogens

Streptococcus pneumoniae Haemophilus influenzae

Klebsiela pneumonia and other


Staphylococcus aureus
Enterobacteriaceae

Moraxella catarrhalis Mycobacterium spp.

Fusobacterium spp Bordetella spp

Pseudomonas spp Legionella spp.

Chlamydia pneumoniae

Criteria of specimen rejection


Saliva (report as “Improper specimen, only saliva, please resubmit)
Expectorated Sputum
All expectorated sputum is contaminated to some degree with
secretion of the oropharngeal cavity, which contain a wide variety of
commensals bacteria, some of which are potential pathogens of the
lower respiratory tract (Strepto. pneumonia, Haemophilus influenzae)
,since the sputum reflect the infection in the branchi and the lung
contamination with oropharyngeal secretion should be kept to a
minimum.

Early morning sputum samples should be obtained because they


contain pooled overnight secretions in which pathogenic bacteria are
more likely to be concentrated, twenty four- hour collection should be
discouraged because there is not only a greater likelihood of
contamination but bacteria pathogens become diluted with the addition
of subsequent, more watery specimens.
Expectorated Sputum continue …….
Instruct the patient to brush his teeth and gargle with water
immediately before obtaining the sputum specimen to reduces the
number of contaminating oro-pharyngeal bacteria

To prevent contaminated of the out side of the container the patient


should be instructed to press the rim of the container and the lower lip
to catch the entire expectorated cough sample.

When a sputum specimen is plated out, it is best to get the portion of


the sample that most looks like pus onto the swab. If there is any
blood in the sputum, this should also be on the swab.
Specimen Processing

Note Trans-tracheal and percutaneous lung aspiration material maybe inoculated


to enrich Thioglycolate and anaerobic blood agar plate.
Sputum’s Gram Stain
Gram stain important to evaluate the quality and realty of the sputum
specimen, an acceptable specimen yield less than 10 sequamous epithelial
cells per low power field (100x).

On the other hand the presence of 25 or more polymorphonuclear


leukocytes per 100x field, together with few sequamous epithelial cells
implies an excellent specimen.
Unacceptable specimen
This low-power (100x) view of a sputum specimen shows many squamous cells, each of
which has a single nucleus surrounded by a large volume of cytoplasm.
Acceptable specimen
Gram’s Stain of Sputum Specimen

Gram stain of sputum specimen ( 400x)


A, this specimen contains numerous polymorphonuclear leukocytes and no visible sequamous epithelial
cells, indicating that the specimen is acceptable for routine bacteriological culture .

B, this specimen contains numerous sequamous epithelial cells and rare polymorphonuclear leukocytes,
indicating an inadequate specimen for routine sputum culture.
Unacceptable specimen
This high-power (400x) view of a sputum specimen shows many squamous cells, each of
which has a single nucleus surrounded by a large volume of cytoplasm.
Note the numerous polymorphonuclear neutrophils and gram-positive, lancet-shaped
diplococci.
ACID FAST STAIN
Ziehl Neelsen method
Initial Processing
Result Reporting of AFS

AFB* Acid Fast Bacilli


Acid Fast bacilli
Post specimen processing
Interfering factors:
Patient on antibiotic therapy.
Improper sample collection.

Result reporting:
Report Gram stain and AFS finding as an initial report.
Report the isolated and its sensitivity pattern as a final report.

Turn around time:


Gram stain and AFS results should be available an 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.

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