Professional Documents
Culture Documents
Chapter 20: Microorganisms Encountered in The Respiratory Tract
Chapter 20: Microorganisms Encountered in The Respiratory Tract
Culture 3. Transport
Routine culture: 5% sheep BA (CO2), MacConkey, CA(CO2)
Multiple interrupted streaking with heating Specimen should be examined without delay or transported within 2
Additional media may be used: hours
SSA (Streptococcal Selective Agar): will supress the normal Refrigeration of specimen for not > 24 hours (2-3 hours) is satisfactory
flora and other beta-hemolytic streptococci except Group A for the recovery of most pathogens
and B; this will hasten the isolation and ID of organisms
4. Laboratory Examination o Children (between 1 mo. – 6yrs. old): H. influenza type
B (most common but has now been reduced because of
First observe macroscopically (describe the gross appearance) newborn screening)
Microscopic examination o Children (6 years old) : meningococcal or
Gram stain
Test of reliability pneumococcal meningitis
Will evaluate the quality of expectorated sputum received for o Can be diagnosed through microscopic examinations,
routine bacterial culture chemical studies and hematology
Purulent culture is carefully selected glass slide air dry o CSF: purulent
stain LPO (100x) Numbers of inflammatory cells (PMN) >
Slide should be scanned 100x and the number of
1000/ cumm – neutrophils
squamous epithelial cells (SEC) and
polymorphonuclear (PMN) leukocyte noted Glucose levels < 45 mg/dL
Ideal specimen: < 10 SEC, > PMN/LPF Protein concentration > 100 mg/dL
Acceptable specimen: < 10 SEC/LPF o May be acute: usually caused by the encapsulated
Exception: patients who are neutropenic bacterial species (ex. H. influenzae & S. pneumoniae)
Contaminated specimen: > 10 SRC/LPF o May be chronic: caused by M. tuberculosis, other
Acid fast stain: classic Ziehl – Neelsen or Kinyoun carbol fuchsin bacteria, fungi (patient would have tuberculous
2 smears each for direct and concentration method meningitis)
Culture
- Tuberculosis meningitidis
Endotracheal aspirates (ETAs) from mechanically ventilated adult
patients can be screened by gram stain o CSF is non purulent
Criteria used to reject ETAs from adult patients included greater o Reduced glucose content
than 10 squamous epithelial cells per LPO field or no organisms o CHON increase (> 50 mg/dL)
seen under OIO o Low white cell count of mononuclear type
Exception: in Legionella pneumonia, sputum may be scanty
and watery, with few or no host cells. Such specimens may
be positive by direct florescent antibody stain and culture, 1. Collection, Transport and Initial Handling of Specimen
and they should not be subjected to screening procedures
Sputum from patients with CF (cystic fibrosis) should be screened. Lumbar tap
A throat swab is an acceptable specimen from patients with CF in Skin must first be decontaminated to avoid normal flora
selected clinical settings and should be processed in a similar Hand delivered immediately to the laboratory (because most are
manner as CF sputum fastidious organisms)
For routine culture: 1-3mL of sputum is needed
Should NEVER be refrigerated
5% sheep BA (CO2), CA (CO2), MacConkey, Thio
Pick out purulent or bloody portions of the specimen for If not processed immediately, incubate at 37C or leave at RT for 6
inoculation hours
For increased recovery of Hi, incorporate bacitracin into CA If specimen is suspected of hatbouring a virus, refrigerate for up to
AFB culture 24 hours after collection or frozen at -7-C
Ziehl – Neelsen or the Kinyuon carbol fuchsin stain Concentrated by centrifugation
Auramine or auramine – rhodamine can also aid in detecting Sediment – Microbiological studies
AFB (more sensitive and more expensive)
Supernatant – Chemical studies
10mL of sputum
2 LJ slants 3 or 4 tubes should be collected
Any typical growth of possible pathogen do ID Tube 1 & 2: Microbiological and chemical studies
Method of streaking (CVGH) for routine culture: multiple Tube 3 & 4: Cell count and differential
interrupted streaking with heating in between areas of streaking We use the last two tubes because usually when the
Purpose: quantitation (BA) doctor collects the first two tubes its still clear while the
Organisms causing inflammation will be present in greater
last two tubes contain blood
numbers than any other superficial contaminating organism
Semiquantitation: Volume: 1-5 mL